WORLD INTELLECTUAL PROPERTY ORGANIZATION
International Bureau
PCT
INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
(51) International Patent Classification 6 :
C07D 263/32, A61K 31/42, C07D 413/12,
C07C 275/28, 275/34, 275/42
Al
(11) International Publication Number: WO 97/40028
(43) International Publication Date: 30 October 1997 (30.10.97)
(21) International Application Number: PCT/US97/06623
(22) International Filing Date: 21 April 1997 (21.04.97)
(30) Priority Data:
08/636,361
08/801,780
08/832,165
23 April 1996 (23.04.96) US
14 February 1997 (14.02.97) US
2 April 1997 (02.04.97) US
(71) Applicant: VERTEX PHARMACEUTICALS INCORPO-
RATED fUS/US); 130 Waverly Street, Cambridge, MA
02139-4242 (US).
(72) Inventors: ARMISTEAD, David, M.; Five Cutting Drive,
Maynard, MA 01574 (US). BADIA, Michael, C; 20
Meadowbrook Road, Bedford, MA 01730 (US). BEMIS,
Guy, W.; 256 Appleton Street, Arlington, MA 02174 (US).
BETHIEL, Randy, S.; 28 Haskell Street. Allston, MA
02134 (US). FRANK, Catharine, A.; 374 Simpson Road,
Marlborough, MA 01752 (US). NOVAK, Perry, M.; 35
Debbie Lane, Milford, MA 01757 (US). RONKIN, Steven,
M.; 39 Bridge Street #14, Watertown, MA 02172 (US).
SAUNDERS, Jeffrey, O.; 164 Parker Street, Acton, MA
01720 (US).
(74) Agents: HALEY, James, F., Jr. et al.; Fish & Neave, 1251
Avenue of the Americas, New York, NY 10020-1 104 (US).
(81) Designated States: AL, AM, AT, AU, AZ, BA, BB, BG, BR,
BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, GE,
GH, HU. IL, IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR,
LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, NO, NZ,
PL, PT, RO, RU, SD, SE, SG, SI, SK, TJ, TM, TR, TT,
UA, UG, UZ, VN, YU, ARIPO patent (GH, KE, LS, MW,
SD, SZ, UG), Eurasian patent (AM, AZ, BY. KG, KZ, MD,
RU, TJ, TM), European patent (AT, BE, CH, DE, DK, ES,
FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OAPI patent
(BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, NE, SN, TD,
TG).
Published
With international search report.
(54) Title: UREA DERIVATIVES AS INHIBITORS OF IMPDH ENZYME
(57) Abstract
The present invention relates to a novel class of compounds which are IMPDH inhibitors. This invention also relates to pharmaceutical
compositions comprising these compounds. The compounds and pharmaceutical compositions of this invention are particularly well suited
for inhibiting IMPDH enzyme activity and consequently, may be advantageously used as therapeutic agents for IMPDH mediated processes.
This invention also relates to methods for inhibiting the activity of IMPDH using the compounds of this invention and related compounds.
FOR THE PURPOSES OF INFORMATION ONLY
Codes used to identify States party to the PCT on the front pages of pamphlets publishing international applications under the PCT.
AL
Albania
ES
Spain
LS
Lesotho
SI
Slovenia
AM
Armenia
FI
Finland
LT
Lithuania
SK
Slovakia
AT
Austria
FR
France
LU
Luxembourg
SN
Senegal
AU
Australia
GA
Gabon
LV
Latvia
sz
Swaziland
AZ
Azerbaijan
GB
United Kingdom
MC
Monaco
TD
Chad
BA
Bosnia and Herzegovina
GE
Georgia
MD
Republic of Moldova
TG
Togo
BB
Barbados
GH
Ghana
MG
Madagascar
TJ
Tajikistan
BE
Belgium
GN
Guinea
MK
The former Yugoslav
TM
Turkmenistan
BF
Burkina Faso
GR
Greece
Republic of Macedonia
TR
Turkey
BC
Bulgaria
HI)
Hungary
ML
Mali
TT
Trinidad and Tobago
BJ
Benin
IE
Ireland
MN
Mongolia
UA
Ukraine
BR
Brazil
IL
Israel
MR
Mauritania
UG
Uganda
BY
Belarus
IS
Iceland
MW
Malawi
US
United States of America
CA
Canada
IT
Italy
MX
Mex ico
UZ
Uzbekistan
CF
Central African Republic
JP
Japan
NE
Niger
VN
Viet Nam
CG
Congo
KK
Kenya
NL
Netherlands
YU
Yugoslavia
CH
Switzerland
KG
Kyrgyzstan
NO
Norway
7.W
Zimbabwe
CI
Cote d'lvoire
KP
Democratic People's
NZ
New Zealand
CM
Cameroon
Republic of Korea
PL
Poland
CN
China
KR
Republic of Korea
PT
Portugal
cu
Cuba
KZ
Kazakstan
RO
Romania
cz
Czech Republic
LC
Saint Lucia
RU
Russian Federation
DE
Germany
LI
Liechtenstein
SD
Sudan
DK
Denmark
I.K
Sri Lanka
SE
Sweden
EE
Estonia
LR
Liberia
SG
Singapore
WO 97/40028
PCTVUS97/06623
UREA DERIVATIVES AS INHIBITORS OF IMPDH ENZYME
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a novel
5 class of compounds which inhibit IMPDH. This invention
also relates to pharmaceutical compositions comprising
these compounds. The compounds and pharmaceutical
compositions of this invention are particularly well
suited for inhibiting IMPDH enzyme activity and
10 consequently, may be advantageously used as therapeutic
agents for IMPDH mediated processes. This invention
aJ.so relates to methods for inhibiting the activity of
IMPDH using the compounds of this invention and related
compounds .
15
BACKGROUND OF THE INVENTION
The synthesis of nucleotides in organisms is
required for the cells in those organisms to divide and
replicate. Nucleotide synthesis in mammals may be
20 achieved through one of two pathways: the de novo
WO 97/40028
PCT/US97/06623
- 2 -
synthesis pathway or the salvage pathway. Different
cell types use these pathways to a different extent.
Inosine-5 ' -monophosphate dehydrogenase
(IMPDH; EC 1.1.1.205) is an enzyme involved in the de
5 novo synthesis of guanosine nucleotides. IMPDH
catalyzes the NAD-dependent oxidation of inosine-5 '-
monophosphate (IMP) to xanthosine- 5 • -monophosphate
(XMP) [Jackson R.C. et. al . , Nature , 256, pp. 331-333,
(1975) ] .
10 IMPDH is ubiquitous in eukaryotes, bacteria
and protozoa [Y. Natsumeda & S.F. Carr, Ann. N.Y.
Acad. , 696, pp. 88-93 (1993)]. The prokaryotic forms
share 30-40% sequence identity with the human enzyme.
Regardless of species, the enzyme follows an ordered
15 Bi-Bi reaction sequence of substrate and cofactor
binding and product release. First, IMP binds to
IMPDH. This is followed by the binding of the cofactor
NAD. The reduced cofactor, NADH, is then released from
the product, followed by the product, XMP [S.F. Carr et
20 al., J. Biol. Chem. , 268, pp. 27286-90 (1993); E.W.
Holmes et al . , Biochim. Biophys. Acta , 364, pp. 209-217
(1974)]. This mechanism differs from that of most
other known NAD-dependent dehydrogenases, which have
either a random order of substrate addition or require
25 NAD to bind before the substrate.
Two isoforms of human IMPDH, designated type
I and type II, have been identified and sequenced [F.R.
Collart and E. Huberman, J. Biol. Chem. , 263, pp.
15769-15772, (1988); Y. Natsumeda et . al., J. Biol.
30 Chem. , 265, pp. 5292-5295, (1990)]. Each is 514 amino
acids, and they share 84% sequence identity. Both
IMPDH type I and type II form active tetramers in
solution, with subunit molecular weights of 56 kDa [Y.
WO 97/40028
PCT/US97/06623
- 3 -
Yamada et . al . , Biochemistry , 27, pp. 2737-2745
(1988) ] .
The de novo synthesis of guanosine
nucleotides, and thus the activity of IMPDH, is
5 particularly important in B and T- lymphocytes . These
cells depend on the de novo, rather than salvage
pathway to generate sufficient levels of nucleotides
necessary to initiate a proliferative response to
mitogen or antigen [A.C. Allison et . al., Lancet II ,
10 1179, (1975) and A.C. Allison et . al . , Ciba Found .
Symp. , 48, 207, (1977)]. Thus, IMPDH is an attractive
target for selectively inhibiting the immune system
without also inhibiting the proliferation of other
eel Is .
15 Immunosuppression has been achieved by
inhibiting a variety of enzymes including for example,
the phosphatase calcineurin (inhibited by cyclosporin
and FK-506) ; dihydroorotate dehydrogenase, an enzyme
involved in the biosynthesis of pyrimidines (inhibited
20 by leflunomide and brequinar) ; the kinase FRAP
(inhibited by rapamycin) ; and the heat shock protein
hsp70 (inhibited by deoxyspergualin) . [See B. D. Kahan,
Immunological Reviews , 136, pp. 29-49 (1993); R. E.
Morris, The Journal of Heart and Lung Transplantation ,
25 12(6), pp. S275-S286 (1993)].
Inhibitors of IMPDH are also known. United
States patents 5,380,879 and 5,444,072 and PCT
publications WO 94/01105 and WO 94/12184 describe
mycophenolic acid (MPA) and some of its derivatives as
30 potent, uncompetitive, reversible inhibitors of human
IMPDH type I (Kj_ = 33 nM) and type II (Kj_ = 9 nM) . MPA has
been demonstrated to block the response of B and T-
WO 97/40028
PCTYUS97/06623
- 4 -
cells to mitogen or antigen [A. C. Allison et. al . ,
Ann. N. Y. Acad. Sci . , 696, 63, (1993) .
Immunosuppressants, such as MPA, are useful
drugs in the treatment of transplant rejection and
autoimmune diseases. [ R . E. Morris, Kidney Intl. , 49,
Suppl. 53, S-26, (1996)]. However, MPA is
10 characterized by undesirable pharmacological
properties, such as gastrointestinal toxicity and poor
bioavailability. [L. M. Shaw, et. al . , Therapeutic Drug
Monitoring , 17, pp. 690-699, (1995) ] .
Nucleoside analogs such as tiazofurin,
15 ribavirin and mizoribine also inhibit IMPDH [L.
Hedstrom, et. al . Biochemistry , 29, pp. 849-854
(1990)]. These compounds, which are competitive
inhibitors of IMPDH, suffer from lack of specificity to
this enzyme.
20 Mycophenolate mofetil, a prodrug which
quickly liberates free MPA in vivo, was recently
approved to prevent acute renal allograft rejection
following kidney transplantation. [L. M. Shaw, et.
al . , Therapeutic Drug Monitoring , 17, pp. 690-699,
25 (1995); H. W. Sollinger, Transplantation , 60, pp. 225-
232 (1995)]. Several clinical observations, however,
limit the therapeutic potential of this drug. [L. M.
Shaw, et. al., Therapeutic Drug Monitoring , 17, pp.
690-699, (1995)]. MPA is rapidly metabolized to the
WO 97/40028
PCT/US97/06623
- 5 -
inactive glucuronide in vivo. [A.C., Allison and E.M.
Eugui, Immunological Reviews , 136, pp. 5-28 (1993)].
The glucuronide then undergoes enterohepatic recycling
causing accumulation of MPA in the gastrointestinal
5 tract where it cannot exert its IMPDH inhibitory
activity on the immune system. This effectively lowers
the drug's in vivo potency, while increasing its
undesirable gastrointestinal side effects.
It is also known that IMPDH plays a role in
10 other metabolic events. Increased IMPDH activity has
been observed in rapidly proliferating human leukemic
cell lines and other tumor cell lines, indicating IMPDH
as a target for anti-cancer as well as
immunosuppressive chemotherapy [M. Nagai et. al . ,
15 Cancer Res . , 51, pp. 3886-3890, (1991)]. IMPDH has
also been shown to play a role in the proliferation of
smooth muscle cells, indicating that inhibitors of
IMPDH, such as MPA or rapamycin, may be useful in
preventing restenosis or other hyperprolif erative
20 vascular diseases [C. R. Gregory et al . ,
Transplantation , 59, pp. 655-61 (1995); PCT publication
WO 94/12184; and PCT publication WO 94/01105].
Additionally, IMPDH has been shown to play a
role in viral replication in some viral cell lines.
25 [S.F. Carr, J. Biol. Chem. , 268, pp. 27286-27290
(1993)]. Analogous to lymphocyte and tumor cell lines,
the implication is that the de novo, rather than the
salvage, pathway is critical in the process of viral
replication .
30 The IMPDH inhibitor ribavirin is currently
being evaluated for the treatment of hepatitis-C virus
(HCV) and hepatitis-B virus (HBV) infection and
disease. Ribavirin enhances the sustained efficacy of
WO 97/40028
PCT/US97/06623
- 6 -
interferon in HBV and HCV treatment. However, the
therapeutic potential of ribavirin is limited by its
lack of a sustained response in monotherapy and broad
cellular toxicity.
Thus, there remains a need for potent IMPDH
inhibitors with improved pharmacological properties.
Such inhibitors would have therapeutic potential as
immunosuppressants, anti-cancer agents, anti-vascular
hyperprolif erative agents, antiinflammatory agents,
antifungal agents, ant ipsoriat ic and anti-viral agents.
0
SUMMARY OF THE INVENTION
The present invention provides compounds, and
pharmaceutically acceptable derivatives thereof, that
.5 are useful as inhibitors of IMPDH. These compounds can
be used alone or in combination with other therapeutic
or prophylactic agents, such as anti-virals, anti-
inflammatory agents, antibiotics, and
immunosuppressants for the treatment or prophylaxis of
transplant rejection and autoimmune disease.
Additionally, these compounds are useful, alone or in
combination with other agents, as therapeutic and
prophylactic agents for antiviral, anti-tumor, anti-
cancer, anti: ^lammatory agents, antifungal agents,
antipsoriatic immunosuppressive chemotherapy and
restenosis therapy regimens.
The invention also provides pharmaceutical
compositions comprising the compounds of this
invention, as well as multi-component compositions
comprising additional IMPDH compounds together with an
immunosuppressant. The invention also provides methods
of using the compounds of this invention, as well as
other related compounds, for the inhibition of IMPDH.
WO 97/40028
PCT/US97/06623
- 7 -
The compounds of this invention, as well as
those used in the methods of this invention demonstrate
a different metabolic profile than MPA and its
derivatives. Because of this difference, methods of
5 this invention and the compounds used therein may offer
advantages as therapeutics for IMPDH mediated disease.
These advantages include increased overall therapeutic
benefit and reduction in deleterious side effects.
10
15
20
25
30
DETAILED DESCRIPTION OF THE INVENTION
In order that the invention herein described
may be more fully understood, the following detailed
description is set forth. In the description, the
following abbreviations are used:
Designation Reagent or Fragment
Ac
Me
Et
Bn
CDI
DIEA
DMAP
DMF
DMSO
EDC
EtOAc
THF
acetyl
methyl
ethyl
benzyl
carbonyldiimidazole
diisopropyl ethyl amine
dime thylaminopyri dine
dimethyl f ormamide
dimethylsulf oxide
1- (3-dimethylaminopropyl ) -3-
ethylcarbodiimide hydrochloride
ethyl acetate
tetrahydrof uran
The following terms are employed herein:
Unless expressly stated to the contrary, the
terms "-SO2-" and "-S(0) 2 -" as used herein refer to a
WO 97/40028 PCT/US97/06623
sulfone or sulfone derivative (i.e., both appended
groups linked to the S), and not a sulfinate ester.
The terms "halo" or "halogen" refer to a
radical of fluorine, chlorine, bromine or iodine.
5 The term "immunosuppressant" refers to a
compound or drug which possesses immune response
inhibitory activity. Examples of such agents include
cyclosporin A, FK506, rapamycin, lef lunomide ,
deoxyspergualin, prednisone, azathioprine ,
10 mycophenolate mofetil, OKT3 , ATAG, interferon and
mizoribine .
The term "interferon" refers to all forms of
interferons, including but not limited to alpha, beta
and gamma forms .
15 iMPDH-mediated disease refers to any disease
state in which the IMPDH enzyme plays a regulatory role
in the metabolic pathway of that disease. Examples of
IMPDH-mediated disease include transplant rejection and
autoimmune diseases, such as rheumatoid arthritis,
20 multiple sclerosis, juvenile diabetes, asthma, and
inflammatory bowel disease, as well as inflammatory
diseases, cancer, viral replication diseases and
vascular diseases.
For example, the compounds, compositions and
25 methods of using them of this invention may be used in
the treatment of transplant rejection (e.g., kidney,
liver, heart, lung, pancreas (islet cells) , bone
marrow, cornea, small bowel and skin allografts and
heart valve xenografts) and autoimmune diseases, such
as rheumatoid arthritis, multiple sclerosis, juvenile
diabetes, asthma, inflammatory bowel disease (Crohn's
disease, ulcerative colitus), lupus, diabetes, mellitus
myasthenia gravis, psoriasis, dermatitis, eczema,
seborrhoea, pulmonary inflammation, eye uveitis,
30
WO 97/40028 PCT/US97/06623
hepatitis, Grave's disease, Hashimoto's thyroiditis,
Behcet's or Sjorgen's syndrome (dry eyes/mouth),
pernicious or immunohaemolytic anaemia, idiopathic
adrenal insufficiency, polyglandular autoimmune
5 syndrome, and glomerulonephritis, scleroderma, lichen
planus, viteligo (depigmentation of the skin) ,
autoimmune thyroiditis, and alveolitis, inflammatory
diseases such as osteoarthritis, acute pancreatitis,
chronic pancreatitis, asthma and adult respiratory
10 distress syndrome, as well as in the treatment of
cancer and tumors, such as solid tumors, lymphomas and
leukemia, vascular diseases, such as restenosis,
stenosis and artherosclerosis , and DNA and RNA viral
replication diseases, such as retroviral diseases, and
15 herpes.
Additionally, IMPDH enzymes are also known to
be present in bacteria and thus may regulate bacterial
growth. As such, the IMPDH- inhibi tor compounds,
compositions and methods described herein may be useful
20 in treatment or prevention of bacterial infection,
alone or in combination with other antibiotic agents.
The term "treating" as used herein refers to
the alleviation of symptoms of a particular disorder in
a patient or the improvement of an ascertainable
25 measurement associated with a particular disorder. As
used herein, the term "patient" refers to a mammal,
including a human.
The term "thiocarbamates" refers to compounds
containing the functional group N-SO2-O.
3 0 The terms " HBV" , "HCV" and "HGV" refer to
hepatitis-B virus, hepatitis-C virus and hepatitis-G
virus, respectively.
WO 97/40028
PCT/US97/06623
- 10 -
According to one embodiment, the invention
provides methods of inhibiting IMPDH activity in a
mammal comprising the step of administering to said
mammal, a compound of formula I:
5
(I)
wherein :
10 A is selected from:
( c l _c 6) -straight or branched alkyl , or (C 2 -
C 6 ) -straight or branched alkenyl or alkynyl; and A
optionally comprises up to 2 subst ituents , wherein:
the first of said substituents , if present,
15 is selected from R 1 or R 3 , and
the second of said substituents , if present,
is R 1 ;
B is a saturated, unsaturated or partially
saturated monocyclic or bicyclic ring system optionally
2 0 comprising up to 4 heteroatoms selected from N, O, or S
and selected from the formulae:
wherein each X is the number of hydrogen atoms
25 necessary to complete proper valence;
WO 97/40028
PCT/US97/06623
- 11 -
and B optionally comprises up to 3 subst ituents ,
wherein :
the first of said substituents , if present,
is selected from R 1 , R 2 , r4 or R 5 ;
5 the second of said substituents, if present,
is selected from R 1 or R 4 , and
the third of said substituents, if present,
is R 1 ; and
D is selected from C(O), C(S), or S{0) 2 ;
10 wherein:
each R 1 is independently selected from 1,2-
methylenedioxy, 1 , 2 -ethylenedioxy , R 6 or (CH 2 ) n -Y;
wherein n is 0, 1 or 2 ; and
Y is selected from halogen, CN, N0 2 , CF 3 , 0CF 3 ,
15 OH, SR6, S(0)R6, S0 2 R 6 , NH 2 , NHR« , N(r6) 2 , Nr6r8 , C OOH,
COOR 6 or OR 6 ;
each R 2 is independently selected from (C 1 -C 4 )-
straight or branched alkyl, or (C 2 - C 4 ) -straight or
branched alkenyl or alkynyl; and each R 2 optionally
20 comprises up to 2 substituents, wherein:
the first of said substituents, if present,
is selected from R 1 , R 4 and R 5 • and
the second of said substituents, if present,
is R 1 ;
- 5 r3 is selected from a monocyclic or a bicyclic
ring system consisting of 5 to 6 members per ring,
wherein said ring system optionally comprises up to 4
heteroatoms selected from N, O, or S, and wherein a CH 2
adjacent to any of said N , 0, or s heteroatoms is
10 optionally substituted with C(O); and each R 3
optionally comprises up to 3 substituents, wherein:
WO 97/40028
PCT/US97/06623
- 12 -
the first of said substituents, if present,
is selected from R 1 , R 2 , R 4 or r5 /
the second of said substituents, if present,
is selected from R 1 or R 4 , and
5 the third of said substituents, if present,
is R 1 ;
each R 4 is independently selected from OR 5 ,
OC(0)R 6 , OC(0)R 5 , 0C{O)0R6, 0C(O)0R 5 , OC(0)N(r6) 2 ,
0P(O)(OR 6 ) 2 , SR 6 , SR5, S(0)R6, S(0)r5, S0 2 r6 , S0 2 R 5 ,
10 S0 2 N(R 6 ) 2 , S0 2 NR 5 R«, S0 3 r6, C(0)r5, C(0)OR5, C (O) ,
C(0)OR 6 , NC(0)C(0)R 6 , NC(0)C(0)R5, NC (O) C (O) OR 6 ,
NC(0)C(0)N(R 6 ) 2 , C(0)N(R6) 2 , C (O) N (OR^ ) r€ ,
C(0)N(OR6)R5 ( C(NOR 6 )R6, C(NOR 6 )r5, N(r6) 2 , Nr6 C (0)r1,
NR6c(0)R 6 , NR6c(0)R5, NR 6 C(0)OR 6 , NR6c(0)OR 5 ,
15 NR6C(0)N(R6) 2 , NR 6 C(0)Nr5r6 ( N R 6 S0 2 R 6 , NR6so 2 R 5 ,
NR 6 S0 2 N(R 6 ) 2 , NR6s0 2 NR 5 R 6 , N(OR 6 )R € , N(OR 6 )R 5 ,
P(O) (OR 6 )N(R 6 ) 2 , and P(0)(OR 6 ) 2 ;
each R 5 is a monocyclic or a bicyclic ring system
consisting of 5 to 6 members per ring, wherein said
20 ring system optionally comprises up to 4 heteroatoms
selected from N, O, or S, and wherein a CH 2 adjacent to
said N, O or S maybe substituted with C (O) ; and each R 5
optionally comprises up to 3 substituents, each of
which, if present, is R 1 ;
25 each R 6 is independently selected from H, (C 1 -C 4 ) -
straight or branched alkyl , or (C 2 -C 4 ) straight or
branched alkenyl ; and
each R 6 optionally comprises a substituent that is R 7 ;
R 7 is a monocyclic or a bicyclic ring system
30 consisting of 5 to 6 members per ring, wherein said
WO 97/40028
PCT/US97/06623
- 13 -
ring system optionally comprises up to 4 heteroatoms
selected from N, O, or S, and wherein a CH 2 adjacent to
said N, O or S maybe substituted with C(0); and each R 7
optionally comprises up to 2 substituents independently
5 chosen from H, (C 1 -C 4 ) -straight or branched alkyl, (C 2 -
C 4 ) straight or branched alkenyl, 1 , 2 -methylenedioxy ,
1 , 2 -ethylenedioxy , or (CH 2 ) n -Z;
wherein n is 0, 1 or 2; and
Z is selected from halogen, CN, N0 2 , CF 3 , OCF 3 ,
0 OH, S (C ± -C 4 ) -alkyl, SO ( C ± -C 4 ) -alkyl , S0 2 ( c± - C 4 ) -alkyl ,
NH 2 , NH(C 1 -C 4 ) -alkyl, N {( C]_ -C 4 ) -alkyl ) 2 , N((C 1 -C 4 )-
alkyDR 8 , COOH, C (0) 0 (C 2 -C 4 ) -alkyl or 0(C r C 4 ) - alkyl;
and
R 8 is an amino protecting group; and
5 wherein any carbon atom in any A, R 2 or R 6 is
optionally replaced by 0, S, SO, S0 2 , NH , or N{C 1 -C 4 )-
alkyl .
The term "substituted" refers to the
replacement of one or more hydrogen radicals in a given
3 structure with a radical selected from a specified
group. When more than one hydrogen radical may be
replaced with a substituent selected from the same
specified group, the substituents may be either the
same or different at every position.
> The term "monocyclic or bicyclic ring system
consisting of 5 to 6 members per ring" refers to 5 or 6
member monocyclic rings and 8, 9 and 10 membered
bicyclic ring structures, wherein each bond in each
ring may be possess any degree of saturation that is
chemically feasible. When such structures contain
substituents, those substituents may be at any position
of the ring system, unless otherwise specified.
WO 97/40028 PCT/US97/06623
10
- 14 -
As specified, such ring systems may
optionally comprise up to 4 heteroatoms selected from
N, O or S. Those heteroatoms may replace any carbon
atoms in these ring systems as long as the resulting
compound is chemically stable.
The term "wherein each X is the number of
hydrogen atoms necessary to complete proper valence"
means that X is 0, 1 or 2 hydrogen atoms, depending
upon the identity of the ring atom to which X is bound
(C, N, 0 or S) , the identity of the two adjacent ring
atoms, and the nature of the bonds between the ring
atom to which X is bound and the two adjacent ring
atoms (single, double or triple bond) . In essence,
this definition is meant to exclude from X any
15 substituents other than hydrogen.
The term "amino protecting group" refers to a
suitable chemical group which may be attached to a
nitrogen atom. The term "protected" refers to when the
designated functional group is attached to a suitable
20 chemical group (protecting group) . Examples of
suitable amino protecting groups and protecting groups
are described in T.W. Greene and P.G.M. Wuts,
Protective Groups in Organic Synthesis, 2d. Ed. , John
Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser
25 and Fieser 's Reagents for Organic Synthesis , John Wiley
and Sons (1994); L. Paquette, ed. Encyclopedia of
Reagents for Organic Synthesis , John Wiley and Sons
(1995) and are exemplified in certain of the specific
compounds used in th 3 invention.
According to another embodiment, the
invention provides methods of inhibiting IMPDH in
mammals by administering a compound of the formula
(II) :
30
WO 97/40028
PCT/US97/06623
- 15 -
H H
(ID
wherein B and D are as defined above.
More preferably, in methods employing the
5 compounds of formulae (I) or (II), component B
comprises from 0 to 2 subs t i tuents . According to an
alternate embodiment, the invention provides methods
for inhibiting IMPDH in a mammal employing compounds
(I) or (II), wherein B comprises at least a single
10 substituent selected from the group defined by R 5 .
Preferably, in this embodiment, B is a monocyclic
aromatic ring containing at least one substituent which
is also a monocyclic aromatic ring.
The present invention also provides compounds
15 which are useful in inhibiting IMPDH. According to one
embodiment, the IMPDH inhibitory compound has the
formula (III) :
(III)
20
wherein A, B and D are as defined above;
E is oxygen or sulfur; and
G and G' are independently selected from R 1 or
hydrogen .
25 According to an alternate embodiment, the
invention provides a compound of the formula (IV):
WO 97/40028
PCT/US97/06623
- 16 -
(IV)
wherein B, D, E, G and G' are defined as above and B'
is a saturated, unsaturated or partially saturated
monocyclic or bicyclic ring system optionally
comprising up to 4 heteroatoms selected from N, O, or S
and selected from the formulae:
wherein each X is the number of hydrogen atoms
necessary to complete proper valence;
and B' optionally comprises up to 3 substituents,
wherein :
the first of said substituents, if present,
is selected from R 1 , R 2 , r4 or R 5^
the second of said substituents, if present,
is selected from R 1 or R 4 , and
the third of said substituents, if present,
is R 1 ; wherein X, R 1 , R 2 , r4 and R 5 are defined as
above .
Excluded from this invention are compounds of
formula (IV) wherein B and B* are simultaneously
unsubstituted phenyl and compounds wherein B is
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- 17 -
unsubstituted phenyl and B' is tri-chloro-, tri-bromo
or tri-iodo phenyl.
Preferably, in compounds of formula (IV), B
and B' are phenyl groups comprising at least one
5 substituent each. These compounds are represented by
formula (V) :
H H
(V)
10 wherein K is selected from R 1 or R 4 ; and J is selected
from R 1 , R 2 or R 4 .
Preferred compounds of formula (V) are those
wherein D is -C(O)-, those wherein E is oxygen; those
wherein J is NR 6 C(0)R 5 or NR 6 C{0)R 6 , preferably
15 NR 6 C(0)R 6 , more preferably N (CH 3 ) C (O) R 6 , and more
preferably N (CH 3 ) C <0) CH 3 ; those wherein K is (CH 2 ) n -Y,
preferably OCH 3 (i.e., n is 0, Y is OR 6 , and R 6 is
CH3) ; and those wherein G is hydrogen. More preferred
compounds of formula (V) are those wherein:
20 E is oxygen
J is NR 6 C(0)R5 or NR 6 C(0)R 6 ;
K is (CH 2 ) n -Y; and
G is hydrogen.
Even more preferred compounds of formula (V) are those
25 wherein:
D is -C(O)-;
E is oxygen;
J is NR 6 C(0)R 6 ;
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- 18 -
K is 0CH 3 ; and
G is hydrogen.
Most preferably in such compounds, J is N ( CH 3 ) C (O) R 6 .
Alternate preferred compounds are those of
5 formula V: wherein J is R 2 , those wherein D is -C{0)-,
those wherein E is oxygen, those wherein J is R 2
A A
substituted with R ' preferably wherein R is
NR 6 C(0)OR 5 or NR 6 C(0)OR 6 , more preferably wherein R 4 i
NR 6 C(0)OR 5 , more preferably wherein R 4 is NHC(0)OR 5 ,
10 and more preferably wherein R 4 is NHC (0)0-3-
tetrahydrofuranyl, those wherein K is (CH 2 ) n -Y,
preferably wherein K is OCH 3 , those wherein G is
hydrogen, and those wherein:
D is -C (0) -;
15 E is oxygen;
K is OCH3; and
G is hydrogen.
Alternatively, other preferred compounds
20 include those of formula VI:
VI
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- 19 -
those compounds of formula VI wherein K is OCH3, and
those compounds of formula VI wherein G is hydrogen.
An alternate embodiment of this invention is
compounds of formula V wherein K is selected from R 1 or
5 R 4 ; and J is selected from R 1 , R 2 , R 4 , and R 9 wherein,
R 1 , R 2 , and R 4 , are as defined above and R 9 is
independently selected from (C 1 -C 4 ) -straight or
branched alkyl , or (C 2 -C 4 ) -straight or branched alkenyl
or alkynyl; and each R 9 optionally comprises up to 2
10 substituents selected from NR 6 C ( O ) OR 1 0 , wherein R6 is
as defined above and R 10 is selected from {C^-C s ) -
straight or branched alkyl optionally comprising up to
two substituents selected from NR 6 R 8 , SR e , S0 2 R 6 ,
-(CH 2 ) n -SR 6 , -(CH 2 ) n -OR6, and OR6 , wherein n, R« and
15 R 8 , are as defined above.
In another embodiment, preferred compounds
are those of formula VII:
wherein K is selected from R 1 and R 4 ; and
A, D, R 1 and R 4 are each independently as defined i
claim 1 .
25
WO 97/40028 PCT7US97/06623
20 -
10
15
More preferred compounds of formula VII are
those wherein D is -0(0)-, those wherein A is a
monocyclic aromatic ring substituted with 1-2
substituents selected from the group consisting of
NR 6 C(0)R 6 , NR 6 C(0)R 5 , CH 2 NR 6 C (O) OR 6 , and CH 2 NR 6 C ( O) OR 5 ,
those wherein A is a monocyclic aromatic ring
substituted with 1-2 substituents selected from the
group consisting of CH 2 NR 6 C (0) OR 6 and CH 2 NR 6 C (O) OR 5 '
those A is a monocyclic aromatic ring substituted with
CH 2 NR 6 C (0) OR 5 ' those wherein A is a monocyclic aromatic
ring substituted with CH 2 NHC (0) OR 5 ' those wherein A is
a monocyclic aromatic ring substituted with CH 2 NHC(0)0-
3-tetrahydrofuryl, those wherein K is (CH 2 ) n -Y, those
wherein K is OCH 3 , and those wherein:
A is a monocyclic aromatic ring substituted
with CH 2 NHC (0) 0-3- tetrahydrof uryl ; and
K is OCH3.
20
Alternatively, other preferred compounds of
this invention include those compounds of formula VIII:
VIII
25
wherein D and K are as defined in claim 1.
Another embodiment is those compounds of
formula IX
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- 21 -
K
H
IX
wherein :
5 D is selected from C(O), C(S) and S(0)2/'
K is selected from R 1 and R 4 ; and
J is selected from R 1 , R 2 , and R 4 .
More preferred compounds of formula IX include
those wherein D is -C(0)-, those wherein J is NR 6 C(0)R 5
or NR 6 C(0)R 6 ' those wherein J is NR 6 C(0)R 6 ' those
wherein J is N ( CH 3 ) C ( O ) R 6 ' those wherein J is
N ( CH3 ) C { 0 ) CH3 r those wherein K is (CH 2 ) n -Y, those
wherein K is OCH3, and those wherein:
K is OCH3; and
J is N(CH 3 ) C (0) CH 3 .
Tables IA, IB and IIB list preferred
individual compounds of the invention and preferred
compounds employed in the compositions and methods of
this invention. Table IIA lists preferred compounds
employed in the methods of this invention.
10
15
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Table I A
H H
#
G
K
A
1
H
H
benzyl
Table IB
H H
#
G
K
B'
2
H
H
3-methoxyphenyl
3
H
H
3-thienyl
4
H
H
3, 4-dif luorophenyl
5
H
H
2, 5-dimethoxyphenyl
6
H
H
3-methylthiophenyl
7
H
H
3-bromophenyl
8
H
H
3-cyanophenyl
9
H
H
3-trif luoromethyl-4-
chlorophenyl
10
H
H
2-methyl-3-chlorophenyl
11
H
H
2 -methoxy- 5-methylphenyl
12
H
H
2-methoxyphenyl
I 13
H
H
3-methoxyphenyl
14
H
H
2, 5-dimethoxyphenyl
15
H
H
3-nitrophenyl
1 6
H
H
4-nitrophenyl
17
H
H
3-methylphenyl
18
H
H
3-trif luorome thy lphenyl
19
H
H
2-trif luoromethylphenyl
20
H
H
3- f luorophenyl
21
H
H
4-phenoxyphenyl
22
H
H
3-chlorophenyl
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- 23 -
#
G
K
B *
TJ
xi
TJ
n
j cnioro i nuoropnenyi
H
TJ
Xl
— 3 TT1 "1 7~i /"\ 1^*4 V* t~~\ \ f 1
j dnnnopnenyi
25
H
H
3- (hydroxymethyl ) phenyl
TJ
XI
TJ
n
3 — ace tyl enyl phenyl
? 7
TJ
LI
n
3— hydroxypheny 1
TJ
n
TJ
xl
3— pyr idinyl
^ n
TJ
xi
TJ
XI
4— pyridinyl
TJ
Jl
TJ
n
2— ( 5 -methyl ) thiazolyl
TJ
n
TJ
n
3, 4— ethylenedioxyphenyl
4 0
H
TJ
XX
j— me cnyi-fj— ni tropnenyl
A i
TJ
n
IJ
3 - tr i f luorome thy 1- 4 —
niLropnenyi
42
H
3-chloro
phenyl
TJ
Xl
^ V"\ 1 r~\ /~i
3 -me thylphenyl
A A
4 S
TJ
n
— ■Fin /— \ v" /~\
j i. ± uuro
phenyl
4 fi
TJ
n
J I -L U.O IT O
3 -me thylphenyl
47
H
1 1
XI
3— carbome thoxyme thylphenyl
4 8
H
n
n
3- carboxye thylphenyl
4 9
ii
n
3— dime thy 1 aminopheny 1
50
H
IT
J- [Z- [Z-
lufc; Liiyj. ^ uioxoidiiyi j prieny±
51
H
H
-j aininocaiDonyipnenyi
53
H
H
3- (3-furanyl) -phenyl
54
1 i
u
n
3— carboxymethylpheny 1
55
H
•J lilts L-liKJJ\y
3— me thylphenyl
56
n
3-ni t rophenyl
57
11
J cm. UIU
3— carbome thoxyme thylphenyl
58
H
n
3 -amino -5— me thylphenyl
59
H
3-methoxy
3-aminophenyl
60
u
n
-> JJI OHIO
3 -me thylphenyl
61
TJ
XX
o Liiior o
j-cnloro-4- (5-
oxazoiyi ) pneny l
62
H
3 — ciilo ro
m i iueLnyipyriuyl )
63
H
3 — fhi 1 nrn
j carDoxyinetnyipnenyi
64
H
*3 — hr om c~\
■~-> XJ X. \J1\.W-)
-5 ni l ropneny i
65
H
3 -hrnmn
j dminopnenyi
66
H
H
3- [ 5- (2-
methylpyrimidinyl ) ] phenyl
67
H
H
3- ( 5-oxazolyl ) phenyl
68
H
3-chloro
2- thienyl
69
H
3-chloro
3- thienyl
71
H
3-chloro
3-methoxy carbamoyl -phenyl
72
H
3-chloro
3-acetamidophenyl
73
H
3-chloro
3-iodophenyl
74
H
3-methyl
phenyl
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- 24 -
ff
G
K
B '
7 5
H
3-methyl
3 -me thyl phenyl
7 6
methyl
3-chloro
3 -me thyl phenyl
77
methyl
H
3-methylphenyl
78
H
3-chloro
3-ni trophenyl
79
H
3-chloro
3-aminophenyl
8 0
H
H
3-
(cyclohexylsulf amoyl ) phenyl
8 1
H
H
3- (methylsuf amoyl ) phenyl
82
H
H
3- (phenylsuf amoyl ) phenyl
83
H
3-methoxy
3 -ben zyloxy carbamoyl -phenyl
84
H
3-methoxy
3-acetamidophenyl
85
H
3-chloro
4- (2-methyl) furanyl
86
H
3-chloro
5- (2-methyl) thienyl
88
H
3-carbomethoxy
3-methylphenyl
89
H
3-carbomethoxy
3-ni trophenyl
91
H
3-chloro
4- (2-nitro) thienyl
92
H
3-chloro
4- ( 2-hydroxyamino ) thienyl
93
H
3-chloro
3- <N-
methyl) trif luoroacetamido-
phenyl
94
H
3-chloro
3- (methylamino ) phenyl
95
H
3-chloro
4- (2-amino) thienyl
96
H
3-methoxy
3- tri f luoroacetamidophenyl
97
H
3-methoxy
3- (N-
methyl ) tri f luoroacetamido-
phenyl
98
H
3-methoxy
3- (3 ' -
picolyloxycarbamoyl ) phenyl
99
H
3-methoxy
3- (phenoxycarbamoyl ) phenyl
100
H
3-methoxy
3-dif luoroacetamidophenyl
101
H
3-
acetoxymethyl
3-methylphenyl
102
H
3-
hydroxyme thyl
3-methylphenyl
104
H
H
3-ni tro-4 - f luorophenyl
105
H
3-methoxy
3- ( aminomethyl ) phenyl
[ • T FA ]
10 6
H
3-methoxy
5- (N-acetoxy) indolinyl
i m
J, u /
H
3-methoxy
3- (N-methyl) ace tamidophenyl
108
H
3-methoxy
3- [ (2-oxo-2- (3,4, 5-tri-
methoxyphenyl ) acetyl )
amino ] phenyl
109
H
3-amino
3-methylphenyl
110
H
3-methoxy
3-benzamidophenyl
111
H
3-methoxy
3 -phenyl ace tamidophenyl
112
H
3-methoxy
3- phenyl ureidophenyl
113
H 1 3-methoxy
3- ( t-butoxycarbamoyl
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- 25 -
#
G
K
B '
methyl ) phenyl
114
H
3-methoxy
3- ( cyclopentylacetamido )
phenyl
115
H
3-methoxy
3-methylphenyl
Table IC
H H
C onion un ri
T
±J
116
lNrlO (U) D L DULyl
117
Hfu^r 1 Ci 1 Pl— -1- — Kn -t- wl
iN^ri3U \\j ) v z. Dutyi
1 T Q
lio
NHC (0) O-methyl
119
NTT-TP 1 C>\ n-nhon ul
120
NHC (0) 0- (S) -3-tetrahydrofuranyl
121
NHC (O) 0-2-picolinyl
122
NHC (O) O- (S) -5-oxazolidinonylmethyl
123
NHC (O) 0-4-carbomethoxyphenyl
124
NHC (0) 0-isobutyl
125
NHC (0) 0-allyl
126
NHC (0) 0-5- ( 1 , 3-dioxanyl )
127
NHC (0) 0-4-acetamidophenyl
128
NHC (0) 0-2-furfuryl
129
NHC (0) 0-2-thiofurfuryl
130
NHC (0) 0-2-methoxyethyl
131
NHC (0) 0-4-tetrahydropyranyl
132
NHC (O) O-cyclohexyl
133
NHC (0) O-cyclopentyl
134
NHC (0) O-2-hydroxyethyl
135
NHC (O) O-cyclohexylmethyl
136
NHC (0) 0- (R, S) -3-tetrahydrofuranyl
137
NHC (0) 0-3-pyridyl
138
NHC (0) O-benzyl
139
NHC (0) 0-3- (tBOC-amino) propyl
140
NHC (0) 0-4-hydroxybutyl
141
NHC (O) 0-5-hydroxypentyl
142
NHC (0) 0- (R, S) -2-pyranyl
143
NHC (0) 0-3- (N-tBOC) -piperidinyl
144
NHC (0) 0- (R) -3- (2-oxo-4, 4-
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- 26 -
Compound
L
dimethyl) furanyl
145
NHC (0) O-3-methylthiopropyl
146
NHC (0) 0-4- [ (2, 2-dimethyl) -1,3-
dioxanyl ] methyl
NHC (0) O-2-di- (hydroxymethyl ) ethyl
148
NHC (0) 0-4- (N-tBOC) -piperidinylme thyl
149
NHC (O) 0-3- (N-tBOC) -piper idinylmethyl
150
NHC (0) O- (dibenzyloxymethyl ) methyl
151
NHC (0) O-di- (hydroxymethyl ) methyl
152
NHC (0) 0-2- (N-tBOC) -piperidinylmethyl
153
NHC (0) O-3-piperidinyl-TFA
154
NHC (0) 0- (R, S) - (2-
tetrahydropyranyl ) methyl
155
NHC (0) O-4-piperidinylmethyl-TFA
156
NHC (0) O- (R, S ) -tetrahydrof uranylmethyl
157
NHC (0) O-3-methylsulfonylpropyl
158
NHC (0) 0-3-piperidinylmethyl-TFA
159
NHC (0) 0-2-piperidinylmethyl-TFA
160
NHC (0) O- (R, S) -3-tetrahydrothiophenyl
161
NHC (0) O- (R, s ) -3-tetrahydrothiopvranvl
162
NHC (0) 0-3-methoxypropyl
Table IIA
5 ' ' *
Q1 H H
#
Q 1
Q 2
B
28
3-methoxy
4-methoxy
3-methylphenyl
32
3-nitro
H
3-methylphenyl
33
4-cyano
H
3-methylphenyl
34
3-methoxy
4 -methoxy
3-bromophenyl
35
3-methoxy
4 -methoxy
2 -methoxy- 5-
chlorophenyl
36
3-methoxy
4-methoxy
3-f luorophenyl
37
3-methoxy
4 -methoxy
3-ethylphenyl
38
3-methoxy
4-methoxy
3 -methyl thiophenyl
52
3-chloro
4 -methoxy
3-ni trophenyl
70
4-cyano
3-chloro
3-methylphenyl
87
1-imidazolyl
H
3-methylphenyl
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- 27 -
90
3-hydroxymethyl
4 -methoxy
3-methylphenyl
103
3- ( t-butoxycarbamoyl
methyl )
H
3- (t-butoxycarbamoyl
methyl ) phenyl
Table I IB
H H
#
Qi
Q3
163
Cl
N (Me) (Ac)
164
OMe
N (Me) (Ac)
165
SMe
CH 2 NHC (0)0- (3s) -
tetrahydrofuranyl
166
S (0) 2 Me
N (Me) (Ac)
167
OMe
N (Me) (Ac)
168
SMe
CH 2 NHC (0) 0- ( 3s) -
tetrahydro furanyl
The compounds of Table IIA correspond to
compounds of formula (II) wherein one of said B
components is phenyl with two substi tuents , Q 1 and Q 2 .
In accordance with formula (II):
Q 1 is selected from R 1 , R 2 , r4 G r R 5 / and
Q 2 is selected from R 1 or R 4 .
The compounds of this invention may contain
one or more asymmetric carbon atoms and thus may occur
as racemates and racemic mixtures, single enantiomers,
diastereomeric mixtures and individual dias tereomers .
All such isomeric forms of these compounds are
expressly included in the present invention. Each
stereogenic carbon may be of the R or S configuration.
Combinations of substituents and variables
envisioned by this invention are only those that result
15
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- 28 -
in the formation of stable compounds. The term
"stable", as used herein, refers to compounds which
possess stability sufficient to allow manufacture and
which maintains the integrity of the compound for a
5 sufficient period of time to be useful for the purposes
detailed herein (e.g., therapeutic or prophylactic
administration to a mammal or for use in affinity
chromatography applications) . Typically, such
compounds are stable at a temperature of 40 °C or less,
10 in the absence of moisture or other chemically reactive
conditions, for at least a week.
As used herein, the compounds of this
invention, including the compounds of formulae I-IX,
are defined to include pharmaceutically acceptable
15 derivatives or prodrugs thereof. A "pharmaceutically
acceptable derivative or prodrug" means any
pharmaceutically acceptable salt, ester, salt of an
ester, or other derivative of a compound of this
invention which, upon administration to a recipient, is
20 capable of providing (directly or indirectly) a
compound of this invention. Particularly favored
derivatives and prodrugs are those that increase the
bioavailability of the compounds of this invention when
such compounds are administered to a mammal (e.g., by
25 allowing an orally administered compound to be more
readily absorbed into the blood) or which enhance
delivery of the parent compound to a biological
compartment (e.g., the brain or lymphatic system)
relative to the parent species. Preferred prodrugs
30 include derivatives where a group which enhances
aqueous solubility or active transport through the gut
membrane is appended to the structure of formulae I-IX.
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- 29 -
Pharmaceutically acceptable salts of the
compounds of this invention include those derived from
pharmaceutically acceptable inorganic and organic acids
and bases. Examples of suitable acid salts include
5 acetate, adipate, alginate, aspartate, benzoate,
benzenesulf onate, bisulfate, butyrate, citrate,
camphorate, camphorsulf onate, cyclopentanepropionate,
digluconate, dodecylsulf ate, ethanesul f onate , formate,
fumarate, glucoheptanoate, glycerophosphate, glycolate,
10 hemisulfate, heptanoate, hexanoate, hydrochloride,
hydrobromide, hydroiodide, 2-hydroxyethanesulf onate,
lactate, maleate, malonate, methanesulf onate, 2-
naphthalenesulfonate, nicotinate, nitrate, oxalate,
palmoate, pectinate, persulfate, 3-phenylpropionate,
15 phosphate, picrate, pivalate, propionate, salicylate,
succinate, sulfate, tartrate, thiocyanate, tosylate and
undecanoate. Other acids, such as oxalic, while not in
themselves pharmaceutically acceptable, may be employed
in the preparation of salts useful as intermediates in
20 obtaining the compounds of the invention and their
pharmaceutically acceptable acid addition salts.
Salts derived from appropriate bases include
alkali metal (e.g., sodium), alkaline earth metal
(e.g., magnesium), ammonium and N- (Ci_4 alkyl) 4 + salts.
25 This invention also envisions the quaternization of any
basic nitrogen-containing groups of the compounds
disclosed herein. Water or oil-soluble or dispersible
products may be obtained by such quaternization.
The compounds of this invention may be
50 synthesized using conventional techniques.
Advantageously, these compounds are conveniently
synthesized from readily available starting materials.
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- 30 -
In general, compounds of formula (I) -{IX) are
conveniently obtained via methods illustrated in
General Synthetic Schemes 1-3.
In General Synthetic Scheme 1 (see below), an
5 X-substituted aniline is reacted with a Y-substituted
phenylisocyanate under standard conditions to give the
desired urea. In this process, X and Y may be one or
more independent substituents (or their suitably
protected variants) as exemplified by the ring
10 substituents listed for compounds of formulae I-IX
above, at any position on the aromatic ring.
WO 97/40028
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- 31
General Synthetic Scheme 1 :
H 2 N.
NCO
CHCI 2 /DMF
Ambient temp.
12 hours
H H
General Synthetic Scheme 2:
3) m-tolyl isocyanate
dichloroethane
ambient temp
overnight
General Synthetic Scheme 3:
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- 32 -
In General Synthetic Scheme 2 (see above) , a
substituted benzaldehyde (here, 2-methoxy-4-ni tro-
substituted) is treated sequentially with
tosylmethylisocyanide, to give the resulting oxazole,
5 then reduced by catalytic hydrogenat ion to give the
desired aniline. Reaction of this aniline with an
isocyanate (here, m-tolylisocyanate) under standard
conditions gives the desired urea.
An alternate synthetic route is illustrated
10 in General Synthetic Scheme 3 (see above) . A
substituted benzaldehyde (here 4-nitro substituted) is
converted to the corresponding oxazolyl aniline as
shown in General Synthetic Scheme 2. This aniline is
treated with a substituted benzoic acid (here, 3-
15 methyl-substituted) and a carboxylic acid activating
agent, such as diphenylphosphoryl azide, under standard
reaction conditions, to give the desired urea.
As can be appreciated by the skilled artisan,
the above synthetic schemes are not intended to
20 comprise a comprehensive list of all means by which the
compounds described and claimed in this application may
be synthesized. Further methods will be evident to
those of ordinary skill in the art. Additionally, the
various synthetic steps described above may be
performed in an alternate sequence or order to give the
desired compounds.
The compounds of this invention may be
modified by appending appropriate functionalities to
enhance selective biological properties. Such
modifications are known in the art and include those
which increase biological penetration into a given
biological compartment (e.g., blood, lymphatic system,
central nervous system) , increase oral availability,
WO 97/40028
PCT7US97/06623
increase solubility to allow administration by
injection, alter metabolism and alter rate of
excretion .
The novel compounds of the present invention
5 are excellent ligands for IMPDH. Accordingly, these
compounds are capable of targeting and inhibiting IMPDH
enzyme. Inhibition can be measured by various methods,
including, for example, IMP dehydrogenase HPLC assays
(measuring enzymatic production of XMP and NADH from
10 IMP and NAD) and IMP dehydrogenase spec trophotomet r ic
assays (measuring enzymatic production of NADH from
NAD). [See C. Montero et al . , Clinica Chimica Acta ,
238, pp. 169-178 (1995)].
Pharmaceutical compositions of this invention
15 comprise a compound of formulae (I), (II) or (VII) or a
pharmaceutically acceptable salt thereof; an additional
agent selected from an immunosuppressant, an anti-
cancer agent, an anti-viral agent, antiinflammatory
agent, antifungal agent, antibiotic, or an anti-
20 vascular hyperprolif eration compound; and any
pharmaceutically acceptable carrier, adjuvant or
vehicle. Alternate compositions of this invention
comprise a compound of formulae (III) -(IX) or a
pharmaceutically acceptable salt thereof; and a
25 pharmaceutically acceptable carrier, adjuvant or
vehicle. Such composition may optionally comprise an
additional agent selected from an immunosuppressant, an
anti-cancer agent, an anti-viral agent,
antiinflammatory agent, antifungal agent, antibiotic,
30 or an anti-vascular hyperprolif eration compound.
The term "pharmaceutically acceptable carrier
or adjuvant" refers to a carrier or adjuvant that may
be administered to a patient, together with a compound
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of this invention, and which does not destroy the
pharmacological activity thereof and is nontoxic when
administered in doses sufficient to deliver a
therapeutic amount of the compound.
5 Pharmaceutical^ acceptable carriers,
adjuvants and vehicles that may be used in the
pharmaceutical compositions of this invention include,
but are not limited to, ion exchangers, alumina,
aluminum stearate, lecithin, self-emulsifying drug
10 delivery systems (SEDDS) such as da-tocopherol
polyethyleneglycol 1000 succinate, surfactants used in
pharmaceutical dosage forms such as Tweens or other
similar polymeric delivery matrices, serum proteins,
such as human serum albumin, buffer substances such as
15 phosphates, glycine, sorbic acid, potassium sorbate,
partial glyceride mixtures of saturated vegetable fatty
acids, water, salts or electrolytes, such as protamine
sulfate, disodium hydrogen phosphate, potassium
hydrogen phosphate, sodium chloride, zinc salts,
20 colloidal silica, magnesium trisilicate, polyvinyl
pyrrolidone, cellulose-based substances, polyethylene
glycol, sodium carboxymethylcellulose , polyacrylates ,
waxes, polyethylene-polyoxypropylene-block polymers,
polyethylene glycol and wool fat. Cyclodextrins such
25 as a-, fi>-, and y-cyclodextrin, or chemically modified
derivatives such as hydroxyalkylcyciodextrins,
including 2- and 3-hydroxypropyl-ft-cyclodextrins , or
other solubilized derivatives may also be
advantageously used to enhance delivery of compounds of
30 formulae I-IX.
The pharmaceutical compositions of this
invention may be administered orally, parenterally , by
inhalation spray, topically, rectally, nasally,
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buccally, vaginally or via an implanted reservoir. We
prefer oral administration or administration by
injection. The pharmaceutical compositions of this
invention may contain any conventional non-toxic
5 pharmaceutically-acceptable carriers, adjuvants or
vehicles. In some cases, the pH of the formulation may
be adjusted with pharmaceutically acceptable acids,
bases or buffers to enhance the stability of the
formulated compound or its delivery form. The term
10 parenteral as used herein includes subcutaneous,
intracutaneous, intravenous, intramuscular, intra-
articular, intraarterial, intrasynovial , intrasternal ,
intrathecal, intralesional and intracranial injection
or infusion techniques.
The pharmaceutical compositions may be in the
form of a sterile injectable preparation, for example,
as a sterile injectable aqueous or oleaginous
suspension. This suspension may be formulated
according to techniques known in the art using suitable
dispersing or wetting agents (such as, for example,
Tween 80) and suspending agents. The sterile
injectable preparation may also be a sterile injectable
solution or suspension in a non-toxic parenterally-
acceptable diluent or solvent, for example, as a
solution in 1 , 3-butanediol . Among the acceptable
vehicles and solvents that may be employed are
mannitol, water, Ringer's solution and isotonic sodium
chloride solution. In addition, sterile, fixed oils
are conventionally employed as a solvent or suspending
medium. For this purpose, any bland fixed oil may be
employed including synthetic mono- or digl ycer ides .
Fatty acids, such as oleic acid and its glyceride
derivatives are useful in the preparation of
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injectables, as are natural pharmaceutically-acceptable
oils, such as olive oil or castor oil, especially in
their polyoxyethylated versions. These oil solutions
or suspensions may also contain a long-chain alcohol
5 diluent or dispersant such as those described in
Pharmacopeia Helvetica, Ph . Helv . , or a similar
alcohol, or carboxymethyl celluose or similar
dispersing agents which are commonly used in the
formulation of pharmaceutically acceptable dosage forms
10 such as emulsions and or suspensions Other commonly
used surfactants such as Tweens or Spans and/or other
similar emulsifying agents or bioavailability enhancers
which are commonly used in the manufacture of
pharmaceutically acceptable solid, liquid, or other
15 dosage forms may also be used for the purposes of
formulation .
The pharmaceutical compositions of this
invention may be orally administered in any orally
acceptable dosage form including, but not limited to,
20 capsules, tablets, emulsions and aqueous suspensions,
dispersions and solutions. In the case of tablets for
oral use, carriers which are commonly used include
lactose and corn starch. Lubricating agents, such as
magnesium stearate, are also typically added. For oral
25 administration in a capsule form, useful diluents
include lactose and dried corn starch. When aqueous
suspensions and/or emulsions are administered orally,
the active ingredient may be suspended or dissolved in
an oily phase is combined with emulsifying and/or
30 suspending agents. If desired, certain sweetening
and/or flavoring and/or coloring agents may be added.
The pharmaceutical compositions of this
invention may also be administered in the form of
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suppositories for rectal administration. These
compositions can be prepared by mixing a compound of
this invention with a suitable non-irritating excipient
which is solid at room temperature but liquid at the
5 rectal temperature and therefore will melt in the
rectum to release the active components. Such
materials include, but are not limited to, cocoa
butter, beeswax and polyethylene glycols.
Topical administration of the pharmaceutical
10 compositions of this invention is especially useful
when the desired treatment involves areas or organs
readily accessible by topical application. For
application topically to the skin, the pharmaceutical
composition should be formulated with a suitable
15 ointment containing the active components suspended or
dissolved in a carrier. Carriers for topical
administration of the compounds of this invention
include, but are not limited to, mineral oil, liquid
petroleum, white petroleum, propylene glycol, polyoxy-
20 ethylene polyoxypropylene compound, emulsifying wax and
water. Alternatively, the pharmaceutical composition
can be formulated with a suitable lotion or cream
containing the active compound suspended or dissolved
in a carrier with suitable emulsifying agents.
25 Suitable carriers include, but are not limited to,
mineral oil, sorbitan monostearate , polysorbate 60,
cetyl esters wax, cetearyl alcohol, 2-oct yldodecanol ,
benzyl alcohol and water. The pharmaceutical
compositions of this invention may also be topically
50 applied to the lower intestinal tract by rectal
suppository formulation or in a suitable enema
formulation. Topicall y- transdermal patches are also
included in this invention.
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The pharmaceutical compositions of this
invention may be administered by nasal aerosol or
inhalation. Such compositions are prepared according
to techniques well-known in the art of pharmaceutical
formulation and may be prepared as solutions in saline,
employing benzyl alcohol or other suitable
preservatives, absorption promoters to enhance
bioavailability, f luorocarbons , and/or other
solubilizing or dispersing agents known in the art.
Dosage levels of between about 0.01 and about
100 mg/kg body weight per day, preferably between about
0.5 and about 75 mg/kg body weight per day of the IMPDH
inhibitory compounds described herein are useful in a
monotherapy and/or in combination therapy for the
15 prevention and treatment of IMPDH mediated disease.
Typically, the pharmaceutical compositions of this
invention will be administered from about 1 to about 5
times per day or alternatively, as a continuous
infusion. Such administration can be used as a chronic
20 or acute therapy. The amount of active ingredient that
may be combined with the carrier materials to produce a
single dosage form will vary depending upon the host
treated and the particular mode of administration. A
typical preparation will contain from about 5% to about
25 95% active compound (w/w) . Preferably, such
preparations contain from about 20% to about 80% active
compound .
When the compositions of this invention
comprise a combination of an IMPDH inhibitor of
^0 formulae (I) -(IX) and one or more additional
therapeutic or prophylactic agents, both the IMPDH
inhibitor and the additional agent should be present at
^ge levels of between about 10 to 100%, and more
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preferably between about 10 to 80% of the dosage
normally administered in a monotherapy regimen. The
additional agents may be administered separately, as
part of a multiple dose regimen, from the compounds of
5 this invention. Alternatively, those agents may be
part of a single dosage form, mixed together with the
compounds of this invention in a single composition.
According to one embodiment, the
pharmaceutical compositions of this invention comprise
10 an additional immunosuppression agent. Examples of
additional immunosuppression agents include, but are
not limited to, cyclosporin A, FK506, rapamycin,
leflunomide, deoxyspergualin, prednisone, azathioprine ,
mycophenolate mofetil, OKT3 , ATAG, interferon and
15 mizoribine.
According to an alternate embodiment, the
pharmaceutical compositions of this invention may
additionally comprise an anti-cancer agent. Examples of
anti-cancer agents include, but are not limited to,
20 cis-platin, actinomycin D, doxorubicin, vincristine,
vinblastine, etoposide, amsacrine, mi toxantrone ,
tenipaside, taxol, colchicine, cyclosporin A,
phenothiazines, interferon and thioxantheres .
According to another alternate embodiment,
25 the pharmaceutical compositions of this invention may
additionally comprise an anti-viral agent. Examples of
anti-viral agents include, but are not limited to,
Cytovene, Ganciclovir, trisodium phosphonof ormate,
Ribavirin, d4T, ddl, AZT, and acyclovir.
10 According to yet another alternate
embodiment, the pharmaceutical compositions of this
invention may additionally comprise an anti-vascular
hyperproliferative agent. Examples of ant i- vascular
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hyperproli f erative agents include, but are not limited
to, HMG Co-A reductase inhibitors such as lovastatin,
thromboxane A2 synthetase inhibitors, eicosapentanoic
acid, ciprostene, trapidil, ACE inhibitors, low
5 molecular weight heparin, mycophenolic acid, rapamycin
and 5- ( 3 ' -pyridinylme thyl ) benzof uran-2 -carboxylate .
Upon improvement of a patient's condition, a
maintenance dose of a compound, composition or
combination of this invention may be administered, if
10 necessary. Subsequently, the dosage or frequency of
administration, or both, may be reduced, as a function
of the symptoms, to a level at which the improved
condition is retained when the symptoms have been
alleviated to the desired level, treatment should
15 cease. Patients may, however, require intermittent
treatment on a long-term basis upon any recurrence of
disease symptoms.
As the skilled artisan will appreciate, lower
or higher doses than those recited above may be
20 required. Specific dosage and treatment regimens for
any particular patient will depend upon a variety of
factors, including the activity of the specific
compound employed, the age, body weight, general health
status, sex, diet, time of administration, rate of
25 excretion, drug combination, the severity and course of
the infection, the patient's disposition to the
infection and the judgment of the treating physician.
In an alternate embodiment, this invention
provides methods of treating or preventing IMPDH
30 mediated disease in a a mammal comprising the step of
administrating to said mammal any of the pharmaceutical
compositions and combinations described above. If the
pharmaceutical composition only comprises the IMPDH
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inhibitor of this invention as the active component,
such methods may additionally comprise the step of
administering to said mammal an agent selected from an
antiinflammatory agent, immunosuppressant, an anti-
5 cancer agent, an anti-viral agent, or an anti-vascular
hyperprolif eration compound. Such additional agent may
be administered to the mammal prior to, concurrently
with, or following the administration of the IMPDH
inhibitor composition.
10 In a preferred embodiment, these methods are
useful in suppressing an immune response in a mammal.
Such methods are useful in treating or preventing
diseases, including, transplant rejection (e.g.,
kidney, liver, heart, lung, pancreas (islet cells),
15 bone marrow, cornea, small bowel and skin allografts
and heart valve xenografts), graft versus host disease,
and autoimmune diseases, such as rheumatoid arthritis,
multiple sclerosis, juvenile diabetes, asthma,
inflammatory bowel disease (Crohn's disease, ulcerative
20 colitus), lupus, diabetes, mellitus myasthenia gravis,
psoriasis, dermatitis, eczema, seborrhoea, pulmonary
inflammation, eye uveitis, hepatitis. Grave's disease,
Hashimoto's thyroiditis, Behcet's or Sjorgen's syndrome
(dry eyes/mouth) , pernicious or immunohaemolytic
25 anaemia, idiopathic adrenal insufficiency,
polyglandular autoimmune syndrome, glomerulonephritis,
scleroderma, lichen planus, viteligo (depigmentation of
the skin), autoimmune thyroiditis, and alveolitis.
These methods comprise the step of
30 administering to the mammal a composition comprising a
compound of any of formulae I-IX and a pharmaceutical^
acceptable adjuvant. In a preferred embodiment, this
particular method comprises the additional step of
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administering to said mammal a composition comprising
an additional immunosuppressant and a pharmaceutically
acceptable adjuvant.
Alternatively, this method comprises the step
5 of administering to said mammal a composition
comprising a compound of formulae I-IX; an additional
immunosuppressive agent and a pharmaceutically
acceptable adjuvant.
In an alternate preferred embodiment, these
10 methods are useful for inhibiting viral replication in
a mammal. Such methods are useful in treating or
preventing, DNA and RNA viral diseases caused by, for
example, HTLV-1 and HTLV-2, HIV-1 and HIV-2,
nasopharyngeal carcinoma virus, HBV, HCV, HGV, yellow
15 fever virus, dengue fever virus, Japanese encephalitis
virus, human papilloma virus, rhinoviruses and Herpes
viruses, such as Epstein-Barr , cytomegaloviruses and
Herpes Simplex, Types 1 and 2, or Type 6. [See, United
States patent 5,380,879].
20 These methods comprise the step of
administering to the mammal a composition comprising a
compound of any of formulae I-IX, and a
pharmaceutically acceptable adjuvant. In a preferred
embodiment, this particular method comprises the
25 additional step of administering to said mammal a
composition comprising an additional anti-viral agent
and a pharmaceutically acceptable adjuvant.
Alternatively, this method comprises the step
of administering to said mammal a composition
30 comprising a compound of formulae I-IX; an additional
anti-viral agent and a pharmaceutically acceptable
adj uvant .
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In another alternate preferred embodiment,
these methods are useful for inhibiting vascular
cellular hyperprolif eration in a mammal. Such methods
are useful in treating or preventing diseases,
5 including, restenosis, stenosis, artherosclerosis and
other hyperprolif erative vascular disease.
These methods comprise the step of
administering to the mammal a composition comprising a
compound of any of formulae I- IX, and a
10 pharmaceutically acceptable adjuvant. In a preferred
embodiment, this particular method comprises the
additional step of administering to said mammal a
composition comprising an additional anti-vascular
hyperprolif erative agent and a pharmaceutically
15 acceptable adjuvant.
Alternatively, this method comprises the step
of administering to said mammal a composition
comprising a compound of formulae I-IX; an additional
anti-vascular hyperprolif erative agent and a
20 pharmaceutically acceptable adjuvant.
In another alternate preferred embodiment,
these methods are useful for inhibiting tumors and
cancer in a mammal. Such methods are useful in
treating or preventing diseases, including, tumors and
25 malignancies, such as lymphoma, leukemia and other
forms of cancer.
These methods comprise the step of
administering to the mammal a composition comprising a
compound of any of formulae I-IX, and a
30 pharmaceutically acceptable adjuvant. In a preferred
embodiment, this particular method comprises the
additional step of administering to said mammal a
composition comprising an additional anti-tumor or
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anti-cancer agent and a pharmaceutical!.;/ acceptable
adj uvant .
Alternatively, this method comprises the ste
of administering to said mammal a composition
5 comprising a compound of formulae I-IX; an additional
anti-tumor or anti-cancer agent and a pharmaceutically
acceptable adjuvant.
In another alternate preferred embodiment,
these methods are useful for inhibiting inflammation
0 and inflammatory diseases in a mammal. Such methods
are useful in treating or preventing diseases,
including, osteoarthritis, acute pancreatitis, chronic
pancreatitis, asthma and adult respiratory distress
syndrome .
5 These methods comprise the step of
administering to the mammal a composition comprising a
compound of any of formulae I-IX, and a
pharmaceutically acceptable adjuvant. In a preferred
embodiment, this particular method comprises the
0 additional step of administering to said mammal a
composition comprising an antiinflammatory agent and a
pharmaceutically acceptable adjuvant.
In order that this invention be more fully
understood, the following examples are set forth.
5 These examples are for the purpose of illustration onl
and are not to be construed as limiting the scope of
the invention in any way.
General Materials and Methods
0 All temperatures are recorded in degrees
Celsius. Thin layer chromatography (TLC) was carried
out using 0.25 mm thick E. Merck silica gel 60 F254
plates and elution with the indicated solvent system.
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Detection of the compounds was carried out by treating
the plate with an appropriate visualizing agent, such
as 10% solution of phosphomolybdic acid in ethanol or a
0.1% solution of ninhydrin in ethanol, followed by
5 heating, and/or by exposure to UV light or iodine
vapors when appropriate. Analytical HPLC was carried
out using a Rainin Mycrosorb-MV, 5yx Cyano reverse phase
column, 3.9mm x 150mm, with a flow rate of 1 . OmL/minut e
and a solvent gradient of 5-100% acetonitrile (0.1%
0 TFA) in water (0.1% TFA) . HPLC retention times were
recorded in minutes. NMR spectral data was acquired
using a Bruker AMX500 in the indicated solvent.
The IMP dehydrogenase HPLC assay follows our
standard conditions for the enzymatic production of XMP
5 and NADH from IMP and NAD, but utilizes high pressure
liquid chromatography on a C18 column with ion pairing
reagents to separate all four components. The extent
of reaction is then determined from the resulting
product peak areas. This assay is particularly useful
0 for determining the inhibition profiles of compounds
which have significant absorbance in the UV-visible
region between 290 and 340 nM.
The reaction mixture typically contains 0.1 M
KPi; pH 8.0, 0 . 1M KC1, 0.5 mM EDTA, 2 mM DTT, and 0.2
5 mM each of IMP and NAD. This solution is incubated at
37°C for 10 minutes. The reaction is started by the
addition of enzyme to a final concentration of 20 to
100 nM, and is allowed to proceed for 10 minutes.
After the allotted time, the reaction is quenched by
0 the addition of mycophenolic acid to a final
concentration of 0.01 mM.
The extent of conversion is monitored by HPLC
using a Rainin Microsorb ODS column C18-200 of
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dimensions 4.6 X 10 mm and a solvent system containing
tetrabutylammonium sulfate (5mM) in 0.1 M KPi pH 6.0
with a 0-30% methanol gradient over 15 minutes. A
similar solvent system has been used previously for the
5 purification of halo-IMP derivatives. [L. C. Antionio
and J. C. Wu, Biochemistry , 33, 1753-1759 (1994).] A
UV-monitor set at 254 nM is used to detect the four
components, and the product peaks are integrated to
determine the extent of conversion of the substrates.
10 For the analysis of inhibitors, the compound
in question is dissolved in DMSO to a final
concentration of 20 mM and added to the initial assay
mixture at the desired concentration in a volume of 2-
5% (v/v) . The reaction is started by the addition of
15 enzyme and after 10 minutes is quenched as above.
After HPLC analysis, the product areas are used to
determine the extent of conversion relative to a
control assay containing only DMSO and no test
compound. IC50 or Ki values are determined from non
20 linear least squares fitting of conversion vs
concentration curves to the tight-binding equations of
Henderson. [P. J. F. Henderson, Biochem. J. , 127, 321
(1972) . ]
We have measured the inhibition constants of
25 each compound against IMPDH using an adaptation of the
method first reported by Magasanik. [B. Magasanik, H.
S. Moyed, and L. B. Gehring J. Biol. Chem. , 226, p. 339
(1957) ] .
Insofar as compounds of formulae I-IX are
30 able to inhibit IMPDH, they are of evident clinical
utility for the treatment of IMPDH mediated disease.
These tests are predictive of the compounds ability to
inhibit IMPDH in vivo.
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Experimental Section
Synthesis of Representative Examples:
5
Example 1
Synthesis of Compound 1
H
H
N
T
N
O.
<J
N'
(1)
10
To a solution of 25mg (156|amole) 4-(5-
oxazolyl ) -aniline in 250|iL CH 2 C1 2 was added 50faL (400
Hmole) of benzyl isocyanate at ambient temperature.
After stirring overnight, 1 was isolated in pure form
by filtration with a 3:1 hexanes/CH 2 Cl 2 rinse in a
yield of 21mg (46%). NMR (500MHz, CDC1 3 ) 8 7.86(s),
7.55(d), 7.38(d), 7.22-7.35(m) , 6.39(s), 5.0(brs),
4.43(s). R f 0.30 (5% MeOH/CH 2 Cl 2 ) .
20
Example 2
Synthesis of Compound 43
Bl
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To a solution of glacial acetic acid (46mL) ,
acetic anhydride (46mL ; 485mmole) and 2-chloro-4-
nitrotoluene (5g, 29.1mmole) at 0 °C was added cone.
5 H2SO4 (6.9mL) in a dropwise fashion. Upon complete
addition, Cr0 3 (8.08g, 80.8mmole) was added portion-
wise over 60 mins. Following an additional 15 mins of
stirring at 0 °C, the reaction mixture was poured over
ice and the resulting precipitate was isolated by
10 filtration, rinsing with cold H2O. Purification by
flash chromatography, eluting with a gradient of 15-50%
EtOAc in hexanes, provided 2 . 02g (24%, 40% based on
recovered starting material) Bl as a white solid. The
1 H NMR was consistent with that of the desired
15 structure.
no 2
B2
Compound Bl was dissolved in 1:1
20 ethanol/water (20mL), treated with cone. H2SO4 (2mL)
and refluxed for 1 hour. Upon cooling to ambient
temperature, the reaction was extracted 3x ' s with
diethyl ether. The ethereal solution was washed twice
with water, dried over Na2SC>4 and concentrated in vacuo
25 to yield a yellow solid. Purified product was obtained
through two recrys tal 1 i zat ions from hot Et 2 0 /hexanes ,
yielding 620mg (47.6%) B2 as a lightly yellowed
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10
15
crystalline solid. The NMR was consistent with that
of the desired structure.
tosylmethyl isocyanide (236mg, 1.2mmol), and powdered
K2CO3 (172mg, 1.2mmole) in methanol (13mL) was heated
at reflux for 90 minutes and then stirred overnight at
ambient temperature. Upon concentration to dryness,
the mixture was partitioned between CH2CI2 and water.
The organics were separated, washed with 0 . 5N HC1,
water and brine and then dried over Na2S04 . The
solvent was removed in vacuo to provide a crude yellow
solid. Purified product B3 was obtained through flash
chromatography, eluting with a gradient of 0-2.5% CH3OH
in CH 2 C1 2 , and recrys tal 1 i zat ion (CH 2 Cl 2 /hexanes ) in a
yield of 3 . 3g (68%) as a light yellow crystalline
solid. The 1 H NMR was consistent with that of the
desired structure.
B3
A mixture of B2 (200mg, 1.2mmol),
B4
25
A solution of B3 <150mg, 0.67mmole) in
ethanol (7.5mL) was treated with SnCl2*2H 2 0 (excess;
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ca. 5 equivalents) and heated at reflux for 30 minutes.
The mixture was cooled to ambient temperature, diluted
with diethyl ether and partitioned with 2N NaOH. The
organics were separated, washed with water and brine,
5 dried over Na 2 S04 and concentrated in vacuo. Purified
product B4 was obtained through flash chromatography,
eluting with a gradient of 0-0.5% CH 3 OH in CH 2 C1 2 / in a
yield of 54mg (41.5%) as a light yellow oil. The 1 H
NMR was consistent with that of the desired structure.
10
H H
(43)
To a solution of 20mg (103^mole) B4 in ImL
15 CH 2 C1 2 was added 20u.L m- tolylisocyanate at ambient
temperature. After stirring overnight, 43 was isolated
in pure form by filtration with an EtOAc/hexanes rinse
in a yield of 25mg (74%) . 1h NMR (500MHz, d 6 -DMSO) 5
9.06 (s), 8.73 (s), 8.50 (s), 7.89 (s), 7.73 (d), 7.67
20 (s), 7.42 (d), 7.31 (s), 7.23 (d) , 7.18 (t), 6.82 (d) ,
2.27 (s) . R f 0.28 (5% MeOH/CH 2 Cl 2 ) .
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Example 3
Synthesis of Compound 56
CI (8.14g, 51%) was prepared from 2-methyl-5-
nitroanisole (10. 0g, 60mmole) in a fashion directly
analogous to the preparation of Bl as described above.
The 1 E NMR was consistent with that of the desired
structure .
A stirred suspension of CI (81.94g, 307mmole)
in dioxane (lOOmL) was treated with concentrated HC1
(20mL) and heated at reflux overnight. Upon cooling to
ambient temperature, the product C2 precipitated as a
light yellow crystalline solid in a yield of 40.65g
(73.1%). The filtrate was concentrated to a volume of
ca. 80mL and a second crop of product crystals was
driven from solution by the addition of hexanes,
yielding 8.91g (16.0%). Both batches were identical by
X H NMR and TLC analysis and were consistent with that
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of the desired material. The total yield of C2 was
49. 56g (89.1%) .
5 C3
A solution of C2 (456mg, 2.51mmole),
tosylmethyl isocyanide (490mg, 2.51mmole) and K 2 C0 3
(347mg, 251mmole) were dissolved in methanol and heated
0 to reflux for 1.5 hours. The product mixture was then
concentrated in vacuo, redissolved in CH2CI2/ washed
with water and brine, dried over Na2SC>4 and again
concentrated in vacuo. Purified product C3 was
obtained through recrystallization (Et20/hexanes ) to
5 yield 375mg (68%). The 1 E NMR was consistent with that
of the desired structure.
20
A solution of C3 (4.214g, 19.1mmole) in EtOAc
(150mL) was treated with 10%Pd/C (1.05g, 25 wt . % of C3)
and subjected to 40psi H 2 (g) (Parr Hydrogenation
Apparatus) overnight. The reaction mixture was
25 filtered and concentrated in vacuo. Pure product C4
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was obtained through flash chromatography, eluting with
a gradient of 30-40% EtOAc/hexane s , in a yield of 3.4g
(93%) . The % NMR was consistent with that of the
desired structure.
To a solution of C4 (25mg, 0 . 131mmole) in
CH2CI2 ( lmL) was added toll isocyanate (25(j.L,
10 0.197mmole) at ambient temperature. After stirring
overnight, 56 was isolated in pure form by filtration
with a CH 2 C1 2 rinse in a yield of 42mg (74%) . 1 E NMR
(500MHz, d 6 -DMSO) 6 8.87 (s), 8.64 (s), 8.37 (s), 7.60
(d) , 7.46 (d), 7.42 (s), 7.33 (s), 7.23 (d) , 7.16-7.19
15 (t), 7.05 (dd), 6.80 (d) , 3.92 (s), 2.28 (s). R f 0.46
(5% MeOH/CH 2 Cl 2 ) .
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Example 4
Synthesis of Compound 59
H h
To a solution of C4 (75mg, 0.394mmole) in
dichloroethane (5mL) was added 3-nitrophenyl isocyanate
(97mg, 0.591mmole) at ambient temperature. After
stirring overnight, Dl was isolated in pure form by
filtration with a CH 2 C1 2 rinse in a yield of 110. 3mg
(79%) . The 1 H NMR was consistent with that of the
desired structure.
H H
To a stirred suspension of Dl (95mg,
0.268mmole) in EtOH (20mL) was added SnCl2*2H 2 0 (302mg,
20 1 . 34mmole ) . The reaction mixture was brought to
reflux, at which time dissolution occurred, for 1.5
hours. The solution was cooled to ambient temperature,
diluted with EtOAc, washed with 2N NaOH and brine,
dried (Na2SC>4) and concentrated in vacuo. Pure product
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59 was obtained through flash chromatography (eluting
with a gradient of 2.5-5% MeOH in CH2CI2)/ followed by
selective crystallization of the desired material from
slightly impure fractions in a yield of 15.7mg (18%).
5 3-H NMR (500MHz, d 6 -DMS0) 6 8.83 (s), 8.44 (s), 8.35
(s), 7.59 (d), 7.48 (d) , 7.40 (s), 6.97-7.04 (dd) ,
6.86-6.92 (t), 6.83 (d) , 6.54 (dd), 6.20 (dd), 5.05 (br
s), 3.92 (s) . R f 0.20 (5% MeOH/CH 2 Cl 2 ) .
0 Example 5
Synthesis of Compound 113
15
A solution of 3-aminobenzylamine (826mg,
6.87mmole) and triethylamine (2.39mL, 17.18mmole) was
treated with di- t-butyldicarbonate (1.50g, 6.87mmole)
and the mixture was stirred at ambient temperature for
20 2 hours. The reaction was then diluted with CH 2 C1 2 ,
washed with NaHC0 3 (aq) , water and brine, dried (Na 2 S0 4 )
and concentrated in vacuo. Pure El was obtained by
flash chromatography, eluting with 25% EtOAc in hexanes
in a yield of 200mg (46%). The 1h NMR was consistent
?5 with that of the desired structure.
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H H O
(113)
A solution of C4 (150mg, 0.789mmole) and 1,1-
5 dicarbonylimidiazole (160mg, 0.986mmole) were combined
in THF (5mL) and stirred for 6 hours at ambient
temperature. The precipitation of imidazole was noted.
To this was then added El (351mg, 1.58mmole) and N,N-
dimethylaminopyridine {97mg, 0.789mmole) and the
10 mixture was refluxed overnight, resulting in a
homogenous solution. Upon cooling to ambient
temperature, the reaction was diluted with EtOAc
(20mL) , washed with KHSO4 ( a <3) / water, and brine, dried
(MgSC>4) and concentrated. Pure 113 was obtained
15 through flash chromatography, eluting with a gradient
of 20-30-35% acetone in hexanes in a yield of 164mg
(47%). 1h NMR (500MHz, d 6 -DMS0) 6 8.90 (s), 8.75 <s),
8.38 (s), 7.60 (d) , 7.51 (s), 7.3-7.46 (m) , 7.21-7.27
(t), 7.05 (dd) , 6.87 (d) , 4.12 (d) , 3.93 (s), 1.44 (s).
20 R f 0.21 (5% MeOH/CH 2 Cl 2 ) .
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Example 6
Synthesis of Compound 70
H H
N. Ji
TXT
(70)
10
15
20
A solution of 3-chloro-4-cyanoaniline (500mg,
7.76mmole) and m-tolylisocyanate (l.OmL, 3.17mmole) in
CH2CI2 (3mL) was stirred overnight at ambient
temperature. The reaction mixture was concentrated and
pure 70 was obtained through MPLC, eluting with 1% MeOH
in CH 2 C1 2 , in a yield of 285mg (31%) . J-H NMR (500MHz,
d 6 -DMSO) 6 9.36 (s), 8.88 (s), 7.94 (s), 7.83 (d) ,
7.44 (d), 7.30 (s), 7.24 (d) , 7.15-7.20 (t), 6.82 (d),
2.29 (s) . R f 0.36 (5% MeOH/CH 2 Cl 2 ) .
Example 7
Synthesis of Compound 108
o
HO^ Ji^ ^ „OMe
OMe
25
Gl
To a solution of 3 , 4 , 5- trimethoxyacetophenone
(9.2g, 43.4 mmol) in pyridine (35mL) was added selenium
dioxide (6.3g, 56.7mmol) and the resulting solution was
heated at reflux overnight. The reaction mixture was
cooled to ambient temperature, filtered through celite
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and concentrated to yield a dark brown oil which was
dissolved into ethyl acetate and washed with 1.0 N HC1
and then with saturated NaHCC>3 . The basic aqueous
layer was diluted with ether and acidified with
5 concentrated HC1 . The layers were separated and the
organic phase was washed with brine and then dried
(Na2SC>4) to give 8.4 g of a dark yellow solid.
Recrystallization of this material from ethyl acetate-
hexane then gave Gl (6.8 g) as a pale yellow solid.
10 The 1 H NMR was consistent with that of the desired
structure .
H H O
A mixture of 59 (64mg, 0.20mmole), Gl (300mg,
1.20mmole) and EDC (300mg, 1.6mmole) in THF ( 5mL) was
stirred overnight at ambient temperature. The reaction
was diluted with EtOAc (150mL) , washed with water,
20 dried (MgSO^g) and concentrated in vacuo. Pure 108 was
obtained through MPLC, eluting with a gradient system
of 0-l%MeOH in CH 2 C1 2 , in a yield of 37 . 4mg (35%).
1 H NMR (500MHz, d 6 -DMS0) 5 9.83 (s), 8.23 (s), 8.18
(s), 7.65 (s), 7.61 (s), 7.35 (d), 7.33 (s), 7.29 (s),
25 7.27 (s), 7.11 (s), 7.06-7.10 (t), 6.94-6.99 (t), 6.52
(d)3.68 (s), 3.63 (s), 3.61(s). R f 0.26 (5%
MeOH/CH 2 Cl2) •
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Example 8
Synthesis of Compound 115
5 (115)
A solution of 59 (300mg, 1 . 58mmole ) and m-
toll isothiocyanate (2.0mL, 14.7mmole) in CH 2 C1 2 (5mL)
was stirred at ambient temperature overnight. To drive
10 the reaction to completion, additional m-toll
isothiocyanate (l.OmL, 7.4mmole) was added and the
mixture was heated to reflux for 3 hours. The reaction
was concentrated In vacuo and 115 was obtained in pure
form through MPLC, eluting with 0-5% EtOAc in CH 2 C1 2 /
15 in a yield of 210mg (39%) . NMR (500MHz, d 6 -DMSO) 5
7.90 (s), 7.89 (s), 7.82 (s), 7.75 (d) , 7.64 (s) 7.44
(s), 7.32-7.37 (t), 7.27 ( s ), 7.13-7.21 (m) , 6.91 (dd) ,
3.98 (s) , 2.40 (s) . R f 0.36 (5% MeOH/CH 2 Cl 2 ) -
2 0 Example 9
Synthesis of Compound 97
II
A solution of nitroaniline (l.Og, 7.13mmole)
in CH 2 C1 2 (25mL) was treated with pyridine (2.9mL,
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36ntmole) and trif luoroacetic anhydride (5mL, 36mmole)
and stirred at ambient temperature for 3 hours. The
reaction was diluted further with CH 2 C1 2 , washed with
IN HC1 and brine, dried (MgSO,}) and concentrated in
5 vacuo to yield II (1.61g, 95%) as a white solid. The
1 H NMR was consistent with that of the desired
structure .
12
10
To a slurry of NaH (60% oil dispersion; 34
mg, 1.42mmole) in THF (lOmL) at 0 °C was added a
solution of II (200mg, 0.85mmole) in THF (lOmL) and the
mixture stirred for 1 hour. To this was added methyl
15 iodide (lOOuL, 1.7mmole) and the mixture was stirred
overnight at ambient temperature. The reaction was
poured into water and extracted with EtOAc. The
organics were separated, dried (MgS0 4 ) and concentrated
in vacuo. Pure 12 was obtained through flash
20 chromatography, eluting with 5% EtOAc in hexanes, in a
yield of 163mg (66%) as a yellow solid. The 1 H NMR was
consistent with that of the desired structure.
13
25
A solution of 12 (163mg, 0.66mmole) in
ethanol ( 5mL) was treated with Pd/C (20mg) and
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subjected to H 2 (1 atm. ) for 3 hours. The reaction was
filtered and concentrated in vacuo to yield 13 (120mg,
84%) as a waxy solid. The 1 H NMR was consistent with
that of the desired structure.
(97)
To a solution of triphosgene (31mg,
10 0.104mmole) in dichloroe thane ( ImL) was added in a
dropwise fashion a solution of B4 (50mg, 0 . 2 60mmole )
and diisopropylethylamine (67mg, 518mmole) in
dichloroethane (5mL) . The reaction mixture was stirred
for an additional 1 hour at ambient temperature,
15 treated with 13 (50mg, 0.230 mmole) and stirred
overnight. The entire reaction mixture was subjected
to flash chromatography, eluting with 1% MeOH in
CH2CI2, to provide pure 97 in a yield of 8mg (7%) . 1h
NMR (500MHz, d 6 -DMS0) 6 9.20 (s), 8.98 (s), 8.39 (s),
20 7.67 (s), 7.63 (d) , 7.48 (s), 7.38-7.45 (m) , 7.04-7.10
(t), 3.95 (s), 3.31 (s) . R f 0.37 (5% MeOH/CH 2 Cl 2 ) .
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Example 10
Synthesis of Compound 111
H H
(111)
5
A solution of 59(50mg, 0.154mmole) and
triethylamine (31mg, 0.308mmole) in DMF ( 0 . 5mL) was
treated in a dropwise fashion with phenylacetyl
chloride (25mg, 0 . 1 69mmole ) and the reaction stirred
10 overnight at ambient temperature. The mixture was
diluted with CH 2 C1 2 , washed with NaHC0 3 (aq) and water,
dried over MgSC>4 and concentrated in vacuo. Pure 111
was isolated by flash chromatography, eluting with 2%
MeOH in CH 2 C1 2 , in a yield of 42mg (62%) . 1 U NMR
15 (500MHz, d 6 -DMS0) 6 10.20(s), 8.90 (s), 8.79 (s), 8.39
<s), 7.88 (s), 7.63 (d), 7.53 (d) , 7.44 (s), 7.25-7.40
(m) , 7.22 (t), 7.14 (d) , 7.05 (dd) , 3.96 (s), 3.66 (s).
R f 0.31 (5% MeOH/CH 2 Cl 2 ) .
Example 11
20 Synthesis of Compound 102
Kl
A solution of 2-methyl-5-nitrobenzoic acid
25 (15g, 82.8mmole) in DMF (75mL) was treated with methyl
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iodide (6.7mL, 1 07 . 64mmole ) followed by powdered K2CO3
(17.2g, 12 4 . 2mmole ) (extreme exotherm) and the
suspension stirred at ambient temperature overnight.
The reaction mixture was partitioned between EtOAc and
5 water, the organics separated and washed with water and
brine, dried (Na2SO^) and concentrated in vacuo to
yield Kl (15.86g, 98%) in pure form as an off-white
solid. The ^-H NMR was consistent with that of the
desired structure.
MeO 2 0
(AcO) 2 HC
K2
K2 (4.09g, 16.2%) was prepared from Kl
(15.86g, 81.3mmole) in a fashion analogous to the
preparation of Bl as described above. The ^-H NMR was
consistent with that of the desired structure.
A solution of K2 (2.5g, 8.03mmole) in dioxane
(lOmL) was treated with cone. HC1 ( 0 . 5mL) and the
mixture was heated to reflux for 2 hours. Additional
cone. HC1 (0.5mL) was added and the reaction refluxed
for 3 hours longer. The mixture was diluted with
EtOAc, washed with water and brine, dried (Na2SC>4) and
concentrated in vacuo. Pure K3 was obtained through
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flash chromatography, elating with a gradient of 20-30-
50% Et20 in hexanes, in a yield of 1.14g (68%). Also
isolated was 215mg (11.8%) of the hydrated aldehyde.
The 1 H NMRs were consistent with that of the desired
5 structures.
K4
10 A solution of K3 (300mg, 1.43mmole) in
benzene ( 5mL) was treated with 1,3-propane diol (114J1L,
1.573mmole) and p-TsOH«H 2 0 (27mg, 0.14mmole) and the
mixture was refluxed with Dean-Stark removal of water
for 4.5 hours. The reaction was cooled to ambient
15 temperature, partitioned between EtOAc and dilute
NaHC03, the organics separated, washed with brine,
dried (Na2SC>4) and concentrated in vacuo. Pure K4 was
obtained through flash chromatography, eluting with a
gradient of 20-25% Et20 in hexanes, in a yield of
20 324mg (84.5%) as an off-white crystalline solid. The
NMR was consistent with that of the desired
structure .
K5
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A solution of K4 (289mg, 1.08mmole) in THF
( 5mL) at 0 °C was treated dropwise with a solution of
DIBAL (1.0M in CH 2 Cl2; 2.7mL, 2.7mmole) and stirred
5 for 40 minutes. The reaction was quenched by addition
of saturated Rochelle's salt solution ( 1 OmL) , diluted
with EtOAc and stirred for 30 minutes. The organics
were collected, washed with brine, dried (Na2SC>4) and
concentrated In vacuo to give 250mg (97%) of K5 as a
10 white crystalline solid. The 1 H NMR was consistent
with that of the desired structure.
OBz
K6
15 A solution of K5 (250mg, l.OSmmole) in CH2CI2
(4mL) at 0 °C was treated with pyridine (llO^iL,
1.37mmole), benzoyl chloride (146^iL, 1.26mmole) and 4-
DMAP (catalytic) , and stirred at ambient temperature
overnight. The reaction mixture was diluted with
20 CH 2 C1 2 , washed with 0 . 5N HC1, water and brine, dried
(Na2SC>4) and concentrated in vacuo. Pure K6 was
obtained through flash chromatography, eluting with 10%
EtOAc in hexanes, in a yield of 340mg (99%) as a white
solid. The 1 H NMR was consistent with that of the
25 desired structure.
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A solution of K6 (326mg, 0.99mmole) in
5 dioxane (7mL) was treated with 2 . ON HC1 ( 5mL) and the
mixture heated at 80 °C overnight. The reaction mixture
was diluted with EtOAc and washed with saturated
NaHC0 3 (aq), water and brine, dried (Na 2 S0 4 ) and
concentrated in vacuo. Pure K7 was obtained through
10 flash chromatography, eluting with 30% Et20 in hexanes,
in a yield of 208mg (77.5%) as a white solid. The
NMR was consistent with that of the desired structure.
A solution of K7 (208mg, 0.729mmole) in MeOH
( 6mL) was treated with K2CO3 (lOlmg, 0.765mmole) and
TosMIC (149mg, 0.765mmole) and the solution heated at
20 60 °C for one hour. The reaction was concentrated in
vacuo, redissolved in CH2CI2 and washed with 1 . ON NaOH
(diluted with saturated NaHCC>3). The aqueous portion
was back-extracted with CH 2 Cl2, the organics combined
and washed with water and brine, dried (Na 2 SC>4) and
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concentrated in vacuo. Pure K8 was obtained through
flash chromatography, eluting with a gradient of 10-
50% acetone in hexanes, in a yield^ of 70mg (44%) . The
1 H NMR was consistent with that of the desired
5 structure.
A solution of K8 (70mg, 0.318) in acetic
anhydride ( 1 . 5mL) and pyridine (l.OmL) was treated with
4-DMAP (catalytic) and stirred at ambient temperature
for 3 hours. The mixture was diluted with CH 2 C1 2 ,
washed with 1 . ON HC1, water and brine, dried (Na2S0 4 )
and concentrated in vacuo to provide K9 in a yield of
82mg (98%) as a pale yellow solid. The ^-H NMR was
consistent with that of the desired structure.
A solution of K9 (80mg, 0.305mmole) in dry
EtOH (4mL) was treated with SnCl2'2H 2 0 (241mg,
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1.07mmole) and the mixture heated at 60 °C for 50
minutes. The reaction was diluted with EtOAc, washed
with saturated NaHC0 3 , water and brine, dried (Na 2 S0 4 )
and concentrated in vacuo. Pure K10 was obtained
through flash chromatography, eluting with a gradient
of 20-30% acetone in hexanes, in a yield of 52mg
(73.4%) as a pale yellow oil. The 1 H NMR was
consistent with that of the desired structure.
AcO
10 Kll
A solution of K10 (52mg, 0.224mmole) in
dichloroethane (2mL) was treated with m-tolyl
isocyanate (43^L, 0.336mmole) and stirred overnight at
ambient temperature. The mixture was diluted with
CH 2 Cl2 : hexanes {2:1), filtered and rinsed with the same
solvent system to provide Kll (67mg, 82%) as a white
solid. The 1 H NMR was consistent with that of the
desired structure.
OH
(102)
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A solution of Kll (33mg, 0.09mmole) in MeOH
(2mL) was treated with 1 . ON NaOH (135 uL, 0.135mmole)
and stirred at ambient temperature for 1.5 hours. The
reaction was neutralized by addition of 1 . ON HC1 (135
5 |_tL) and concentrated in vacuo. The white solid was
rinsed with water and CH2CI2 : hexanes (2:1) and dried in
vacuo to provide 102 (20mg, 68%) as a white solid. 1 H
NMR (500MHz, d 6 -DMSO) 8 9.29 (s), 9.00 (s), 8.42 (s),
7.69 (s), 7.55 (m) , 7.37 (s), 7.33 (s), 7.27 (d), 7.16
10 (t), 6.80 (d) , 5.39 (t), 4.58 (s), 2.28 (s). R f 0.13
(1:1 hexanes/acetone) .
Example 12
Synthesis of Compound 106
15
o
(106)
A solution of C4 (50mg, 0 . 2 63mmole ) in THF
(2mL) was treated with CDI (53mg, 0.330mmole) and
stirred at ambient temperature for 4 hours. To this
was added 1 -acetyl- 6-aminoindole (93mg, 0.526mmole,
Sigma Chemical Co.) and 4-DMAP (35mg, 0.289mmole) and
the mixture refluxed overnight. Diluted with EtOAc
(lOOmL), washed with 5% KHSO4, water and brine, dried
(Na2SC>4) and concentrated in vacuo. Redissolved in
EtOAc and filtered to removed insoluble materials and
20
25
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reconcentrated in vacuo. Pure 106 was obtained through
flash chromatography, eluting with a gradient of 50-
60% acetone in hexanes, in a yield of 37mg (36%) as a
white solid. 1 H NMR (500MHz, d 6 -DMSO) 5 8.79 (s), 8.74
5 (s), 8.37 (s), 8.11 (s), 7.62 (d) , 7.47 (s), 7.43 (s),
7.30 (d), 7.13 (d) , 7.14 (d), 4.11 (t), 3.94 (s), 3.07
(t), 2.17 (s) . R f 0.14 (1:1 hexanes/acetone) .
Example 13
(168)
A suspension of 113 (from Example 5) (250mg, 5.76mmol)
in CH 2 Cl2 (lmL) was treated in a dropwise fashion at
ambient temperature with several equivalents of
trif luoroacetic acid and stirred for 90min. The
resulting solution was stripped in vacuo and tritrated
with CH2CI2 and methanol. Pure product 168 was
isolated by filtration in a yield of 258mg (99%). The
1 H NMR was consistent with that of the desired product.
(120)
A suspension of 168 (250mg, 0.55mmol) in 21mL of
CH2CI2/DMF (20:1 by volume) was treated with triethyl
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amine (193pL, 1.38mmol) and stirred at ambient
temperature until homogeneity was reached. The
solution was cooled to 0 C, treated with (S) 3-
tetrahydrof uranyl-A7-oxysuccinimidyl carbonate (635mg,
5 0.608mmol) and allowed to stir overnight with warming
to ambient temperature. The mixture was poured into
ethyl acetate (500mL), washed with NaHC0 3 (aq) ( 2x),
water (2x), and brine (lx), dried over Na2SC>4 and
stripped in vacuo. Pure product 120 was isolated by
10 tritration (30mL CH 2 C1 2 , 100mL ether) in a yield of
212mg (85%). The 1 U NMR was consistent with that of
the desired product.
Example 14
IMPDH Activity Inhibition Assay
We measured the inhibition constants of the
compounds listed in Table III utilizing the following
protocol :
IMP dehydrogenase activity was assayed
following an adaptation of the method first reported by
Magasanik. [Magasanik, B. Moyed, H. S. and Gehring L.
B. (1957) J. Biol. Chem. 226, 339]. Enzyme activity
was measured spectrophotometrically , by monitoring the
increase in absorbance at 340 nm due to the formation
of NADH (e340 is 6220 M" 1 cm - !). The reaction mixture
contained 0.1 M Tris pH 8.0, 0.1 M KC1, 3 mM EDTA, 2 mM
DTT, 0.1 M IMP and enzyme (IMPDH human type II) at a
concentration of 15 to 50 nM. This solution is
incubated at 37°C for 10 minutes. The reaction is
started by adding NAD to a final concentration of 0 . 1M
and the initial rate is measured by following the
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linear increase in absorbance at 340 nm for 10 minutes.
For reading in a standard spectrophotometer (path
length 1 cm) the final volume in the cuvette is 1.0 ml.
The assay has also been adapted to a 96 well microtiter
5 plate format; in this case the concentrations of all
the reagents remain the same and the final volume is
decreased to 200 ul .
For the analysis of inhibitors, the compound in
question is dissolved in DMSO to a final concentration
10 of 20 mM and added to the initial assay mixture for
preincubation with the enzyme at a final volume of 2-
5% (v/v) . The reaction is started by the addition of
NAD, and the initial rates measured as above. Kj_
determinations are made by measuring the initial
15 velocities in the presence of varying amounts of
inhibitor and fitting the data using the tight-binding
equations of Henderson {Henderson, P. J. F. (1972)
Biochem. J. 127 , 321].
These results are shown in Table III. K-l
20 values are expressed in nM. Category "A" indicates
0.01 to 50 nm activity, category "B" indicates 51-1000
nm activity, category "C" indicates 1001 to 10,000 nm
activity, category M D" indicates greater than 10,000 nm
activity. The designation "ND" is used where a given
2 5 compound was not tested.
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Table III
5
Cmpd
K i
Cmpd
Cmpd
J-
u
(nM)
(nM)
w
(nM
c
4 0
p
1 R
o
2
c
4 1
c
1 Q
3
B
4 2
R
I_j
R O
O U
p
4
D
4 3
Q
R 1
c
5
c
4 4
Q O
C
6
c
p
D "3
O O
r!
7
B
r_»
o
8
c
4 7
D
o 3
9
c
4 8
c
R £
L.
10
c
4 9
r
R ~7
JJ
11
c
50
D
R R
O D
p
12
c
51
R Q
p
13
c
52
p
qn
p
14
c
53
Q 1
p
15
c
54
p
Q 9
p
16
c
55
Q "3
-3 -J
7\
17
B
56
B
QZ1
■R
18
c
57
B
P
19
c
58
ID
20
c
59
A
7\
^1
21
c
60
B
ID
22
c
61
D
Q Q
7\
23
c
62
c
inn
1 u u
P
13
24
B
63
c
i n i
j. u ±
p
25
c
64
B
2 6
p
ft R
D O
fcs
103
c
27
c
66
C
104
c
28
c
67
c
105
B
29
D
68
B
106
B
30
C
69
B
107
A
31
D
70
C
108
B
32
D
71
C
109
B
33
D
72
C
110
B
34
C
73
B
111
A
35
C
74
B
112
B
36
C
75
B
113
A
37
C
76
C
114
B
38
D
77
B
115
B
39
D
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Cmpd
K-i
Cmpd
l
#
\ 1 LTl 1
#
\ ITLYl J
lift
X 1 u
n
i. ^
7\
>i
117
a
o
i ^ n
1 JU
7\
X -L vJ
p
X w> X
119
X -J ^_
A
X\
120
A
133
A
121
A
134
A
122
A
135
A
123
A
136
A
124
A
137
B
125
A
138
A
126
A
139
B
127
A
140
A
128
A
141
A
Cmpd
K-i
Cmpd
K-i
#
( mxi j
#
( n_M )
1 / o
14Z
7\
Ijj
7\
/I
X ft O
D
1 JO
7\
/\
_L T T
a
x O /
LJ
145
A
158
B
146
A
159
A
147
A
160
A
148
A
161
A
149
A
162
A
150
A
163
B
151
B
164
B
152
B
165
A
153
A
166
D
154
A
167
B
168
B
Example 15
Anti-Viral Assays
5
The anti-viral efficacy of compounds may be
evaluated in various in vitro and in vivo assays. For
example, compounds may be tested in in vitro viral
replication assays. In. vitro assays may employ whole
10 cells or isolated cellular components. In vivo assays
include animal models for viral diseases. Examples of
such animal models include, but are not limited to,
rodent models for HBV or HCV infection, the Woodchuck
model for HBV infection, and chimpanzee model for HCV
15 infection.
While we have described a number of
embodiments of this invention, it is apparent that our
basic constructions may be altered to provide other
embodiments which utilize the products and methods of
20 this invention. Therefore, it will be appreciated that
the scope of this invention is to be defined by the
appended claims, rather than by the specific
WO 97/40028
PCT/US97/06623
- 75 -
embodiments which have been presented by way of
example .
WO 97/40028
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CLAIMS
10
15
20
We claim:
1. A method of inhibiting IMPDH activity in
a mammal comprising the step of administering to said
mammal a compound of the formula:
N
H
"NT
H
wherein :
A is selected from:
< c l -c 6> -straight or branched alkyl, or (c 2 ~
C 6 ) -straight or branched alkenyl or alkynyl; and A
optionally comprises up to 2 substituents, wherein:
the first of said substituents, if present,
is selected from R 1 or R 3 , and
the second of said substituents , if present,
is R 1 ;
B is a saturated, unsaturated or partially
saturated monocyclic or bicyclic ring system optionally
comprising up to 4 heteroatoms selected from N, O, or S
and selected from the formulae:
or
wherein each X is the number of hydrogen atoms
25 necessary to complete proper valence;
WO 97/40028
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- 77 -
and B optionally comprises up to 3 subs t i tuents ,
wherein :
the first of said substi tuents , if present,
is selected from R 1 , R 2 , r4 or R 5^
5 the second of said substituents, if present,
is selected from R^ or R^ , and
the third of said substituents, if present,
is R 1 ; and
D is selected from C{0), C(S), or S{0)2;
10 wherein:
each R 1 is independently selected from 1,2-
methylenedioxy, 1 , 2-ethylenedioxy , R 6 or (CH 2 ) n -Y;
wherein n is 0, 1 or 2; and
Y is selected from halogen, CN, N0 2 , CF3 , OCF 3 ,
15 OH, SR 6 , S(0)R 6 , S0 2 R 6 , NH 2 , NHR 6 , N(R6) 2 , NR 6 r8, COOH,
COOR 6 or OR 6 ;
each R 2 is independently selected from (C^-C^)-
straight or branched alkyl, or (C 2 -C 4 ) -straight or
branched alkenyl or alkynyl; and each R 2 optionally
20 comprises up to 2 substituents, wherein:
the first of said substituents, if present,
is selected from r! , r4 anc i p>5, and
the second of said substituents, if present,
is R 1 ;
25 r3 is selected from a monocyclic or a bicyclic
ring system consisting of 5 to 6 members per ring,
wherein said ring system optionally comprises up to 4
heteroatoms selected from N, O, or S, and wherein a CH 2
adjacent to any of said N, O, or S heteroatoms is
WO 97/40028
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- 78 -
optionally substituted with C(0); and each R 3
optionally comprises up to 3 substituents, wherein:
the first of said substituents, if present,
is selected from R 1 , R 2 , r4 or R 5^
5 the second of said substituents, if present,
is selected from R 1 or R 4 , and
the third of said substituents, if present,
is R 1 ;
each R 4 is independently selected from OR 5 ,
10 OC(0)R 6 , OC(0)R 5 , OC(0)OR 6 , OC(0)OR 5 , OC(0)N(R 6 ) 2 ,
OP(0)(OR 6 ) 2 , SR 6 , SR 5 , S(0)R 6 , S(0)R 5 , S0 2 R 6 , S0 2 R 5 ,
S0 2 N(R 6 ) 2 , S0 2 NR 5 R 6 , SO3R 6 , C(0)R 5 , C (O) OR 5 , C(0)R 6 ,
C(0)OR 6 , NC(0)C(O)R 6 , NC(0)C(0)R 5 , NC { O ) C ( O) OR 6 ,
NC (O) C (O)N(R 6 ) 2 , C(0)N(R 6 ) 2 , C ( 0) N ( OR 6 ) R 6 ,
15 C (0)N{OR 6 )R 5 , C(NOR 5 )R 6 , C(NOR 6 )R 5 , N(R 6 ) 2 , NR 6 C ( O ) R^ ,
NR 6 C(0)R 6 , NR 6 C(0)R 5 , NR 6 C(0)OR 6 , NR 6 C(0)OR 5 ,
NR 6 C(0)N(R 6 ) 2 , NR 6 C (0)NR 5 R 6 , NR 6 S0 2 R 6 , NR 6 S0 2 R 5 ,
NR 6 S0 2 N (R 6 ) 2 , NR 6 S0 2 NR 5 R 6 , N(OR 6 )R 6 , N(OR 6 )R 5 ,
P (0) (OR 6 ) N(R 6 ) 2 , and P (O) (OR 6 ) 2 ;
20 each R 5 is a monocyclic or a bicyclic ring system
consisting of 5 to 6 members per ring, wherein said
ring system optionally comprises up to A heteroatoms
selected from N, O, or S, and wherein a CH 2 adjacent to
said N, O or S maybe substituted with C(O); and each P 5
25 optionally comprises up to 3 substituents, each of
which, if present, is R 1 ;
WO 97/40028
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each R 6 is independently selected from H, (C1-C4)-
straight or branched alkyl, or (C2-C4) straight or
branched alkenyl; and
each optionally comprises a substituent that is R 7 ;
5 R 7 is a monocyclic or a bicyclic ring system
consisting of 5 to 6 members per ring, wherein said
ring system optionally comprises up to 4 heteroatoms
selected from N, O, or S, and wherein a CH2 adjacent to
said N, O or S maybe substituted with C (O) ; and each R 7
10 optionally comprises up to 2 substituents independently
chosen from H, (C1-C4 ) -straight or branched alkyl, or
(C2-C4) straight or branched alkenyl, 1,2-
methylenedioxy, 1 , 2-ethylenedioxy , or (CH 2 ) n -Z;
wherein n is 0, 1 or 2; and
15 Z is selected from halogen, CN, N0 2 , CF3, OCF3,
OH, S (C 1 -C 4 ) -alkyl, SO (C 1 -C 4 ) -alkyl, S0 2 (C 1 -C 4 ) -alkyl ,
NH 2 , NH (C]_-C 4 ) - alkyl, N ( ( C ]_-C 4 ) - alkyl ) 2 / N((C 1 -C 4 )-
alkyl)R 8 , COOH, C (O) O (Ci-C 4 ) -alkyl or O (C;l-C 4 ) -alkyl;
and
20 R 8 is an amino protecting group; and
wherein any carbon atom in any A, R 2 or R 6 is
optionally replaced by O, S, SO, S0 2 , NH, or N(C 1 -C 4 )-
alkyl.
25 2 - The method according to claim 1, wherein
in said compound, B has from 0 to 2 substituents.
3. The method according to claim 1 or 2,
wherein in said compound, B comprises at least a first
30 substituent and wherein said first substituent is R 5 .
WO 97/40028
PCT/US97/06623
10 -
4. The method according to claim 3, wherein
in said compound, B is a monocyclic aromatic ring and
said first substituent of B is a monocyclic aromatic
5 ring.
5. A method of inhibiting IMPDH activity in
a mammal comprising the step of administering to said
mammal a compound of the formula:
10
N N
H H
wherein :
D and each B are defined as in claim 1
15 6. The method according to claim 5, wherein
in said compound, at least one B has from 0 to 2
substi tuents .
7. The method according to claim 5 or 6,
20 wherein in said compound, one B comprises at least a
first substituent and wherein said first substituent is
8. The method according to claim 7, wherein
25 in said compound, said B is a monocyclic aromatic ring
and said first substituent of said B is a monocyclic
aromatic ring.
9. A compound of the formula:
30 wherein:
WO 97/40028
PCT/US97/06623
- 81 -
A, D, and B are as defined in claim 1/
E is 0 or S; and
G and G' are independently selected from R 1 or H.
10. A compound of the formula
wherein :
10 B and D are as defined in claim 5;
E, G and G* are as defined in claim 9;
B' is a saturated, unsaturated or partially
saturated monocyclic or bicyclic ring system optionally
comprising up to 4 heteroatoms selected from N, O, or S
15 and selected from the formulae:
or
and B ' optionally comprises up to 3 substituents,
20 wherein:
WO 97/40028
PCT7US97/06623
- 82 -
the first of said subst i tuents , if present,
is selected from R 1 , R 2 , R 4 or R 5 ,
the second of said substituents, if present,
is selected from Rl or R 4 , and
5 the third of said substituents, if present,
is R 1 ;
wherein X, Rl, R2, r4
and R- 3 are as defined in claim 5;
wherein if B is unsubstituted phenyl and all of said
substituents present are on B' are R 1 , then at least
10 one of said R 1 substituents is not chloro, bromo or
iodo; and wherein B and B 1 are not simultaneously
unsubstituted phenyl.
11. The compound according to claim 10
15 having the formula:
20
wherein :
K is selected from R 1 and R 4 ; and
J is selected from R-^-, R 2 , and
12. The compound according to claim 11,
wherein D is -C(O)-.
13. The compound according to claim 11,
25 wherein E is oxygen.
14. The compound according to claim 11,
wherein J is NR 6 C(0)R 5 or NR 6 C(0)R 6 .
WO 97/40028
PCT/US97/06623
- 83 -
15. The compound according to claim 14,
wherein J is NR 6 C(0)R 6 .
5 16. The compound according to claim 15,
wherein J is N (CH 3 ) C (O) R 6 .
17. The compound according to claim 11,
wherein K is (CH2) n -Y.
10
18. The compound according to claim 17,
wherein K is OCH3 .
19. The compound according to claim 11,
15 wherein G is hydrogen.
20. The compound according to claim 16,
wherein :
D is -C (0) -;
20 E is oxygen;
K is OCH 3 ; and
G is hydrogen.
21. The compound according to claim 11,
25 wherein J is R 2 .
22. The compound according to claim 21,
wherein E is oxygen.
30 23. The compound according to claim 21,
2 4
wherein J is R substituted with R .
WO 97/40028
PCT/US97/06623
- 84 -
24. The compound according to claim 23,
wherein R 4 is NR 6 C(0)OR 5 or NR 6 C(0)OR 6 .
25. The compound according to claim 21,
5 wherein K is (CH2) n -Y.
26. The compound according to claim 25,
wherein K is OCH3 .
27. The compound according to claim 21,
D is -C(0)-;
E is oxygen;
K is OCH3; and
G is hydrogen.
10
wherein
15
28. A compound of the formula:
wherein K is selected from R 1 and R 4 ; and
A, D, R 1 and R 4 are each independently as defined in
claim 1 .
25 29. The compound according to claim 28,
wherein D is -C(O)-.
WO 97/40028
PCT/US97/06623
- 85 -
30. The compound according to claim 28,
wherein A is a monocyclic aromatic ring substituted
with 1-2 substituents selected from the group
consisting of NR 6 C(0)R 6 , NR 6 C(0)R 5 , CH 2 NR 6 C ( O ) OR 6 , and
5 CH 2 NR 6 C (O) OR 5 .
31. The compound according to claim 30,
wherein A is a monocyclic aromatic ring substituted
with 1-2 substituents selected from the group
10 consisting of CH 2 NR 6 C ( O) OR 6 and CH 2 NR 6 C (O) OR 5 .
32. The compound according to claim 28,
wherein K is (CH 2 ) n -Y.
15
33. The compound according to claim 32,
wherein K is OCH3 .
34. A compound according to the formula:
K
CN
J
U
H
N
H
20
wherein :
D is selected from C(O), C(S) and S(0) 2 ;
K is selected from R 1 and R 4 ; and
J is selected from R 1 , R 2 , anc i r4 .
35. The compound according to claim 34,
wherein D is -C(O)-.
WO 97/40028
PCT/US97/06623
- 86 -
36. The compound according to claim 34,
wherein J is NR 6 C(0)R 5 or NR 6 C(0)R 6 .
5 37. The compound according to claim 34,
wherein K is (CH 2 ) n -Y.
38. The compound according to claim 37,
wherein K is OCH3 .
10
39. The compound according to claim 10
selected from the group consisting of compounds 1-27,
29-31, 39-51, 53-69, 71-86, 88-89, 91-102 and 104-162
in Tables IA, IB and IC.
15
40. The compound according to claim 28
selected from the group consisting of compounds 163-168
in Table IIB.
20
41. A pharmaceutical composition comprising
a. a compound of the formula:
N N
H H
or
N N
H H
25 in an amount effective to inhibit IMPDH activity,
wherein A, B and D are as defined in claim 1 ;
b. an additional agent selected from
an immunosuppressant, an anti-cancer agent, an anti-
viral agent, antiinflammatory agent, antifungal agent,
WO 97/40028 PCT/US97/06623
- 87 -
antibiotic, or an anti-vascular hyperproliferation
agent;
c. a pharmaceutically acceptable
adjuvant .
42. The composition according to claim 41,
wherein in said compound, at least one B comprises at
least a first substituent and wherein said first
substituent is R- 1 .
10
43. A pharmaceutical composition comprising:
a. a compound according to any one of
claims 9 to 40 in an amount effective to inhibit IMPDH
activity; and
15 b. a pharmaceutically acceptable
ad j uvant .
44. The pharmaceutical composition according
to claim 43, additionally comprising an additional
20 agent selected from an immunosuppressant, an anti-
cancer agent, an anti-viral agent, antiinflammatory
agent, antifungal agent, antibiotic, or an anti-
vascular hyperproliferation agent.
25 45. A method for treating or preventing
IMPDH mediated disease in a mammal comprising the step
of administering to said mammal a composition according
to claim 4 1 .
46. A method for treating or preventing
IMPDH mediated disease in a mammal comprising the step
of administering to said mammal a composition according
to claim 43.
WO 97/40028
PCT/US97/06623
- 88 -
47. The method according to claim 46,
wherein said composition additionally comprises an
agent selected from an immunosuppressant, an anti-
5 cancer agent, an anti-viral agent, antiinflammatory
agent, antifungal agent, antibiotic, or an anti-
vascular hyperprolif eration agent
48. The method according to any one of claims
10 45 to 47, wherein said method is used to suppress an
immune response and wherein said additional agent, if
present, is an immunosuppressant.
49. The method according to claim 48,
15 wherein said IMPDH mediated disease is an autoimmune
disease .
50. The method according to any one of
claims 45 to 47, wherein the IMPDH mediated disease is
20 a viral disease and wherein said additional agent, if
present, is an anti-viral agent.
51. The method according to any one of
claims 45 to 47, wherein the IMPDH mediated disease is
25 a vascular disease and wherein said additional agent,
if present, is an anti-vascular hyperproli f eration
agent .
52. The method according to any one of
30 claims 45 to 47, wherein the IMPDH mediated disease is
cancer and wherein said additional agent, if present,
is an anti-cancer agent.
WO 97/40028
PCT/US97/06623
- 89 -
53. The method according to any one of claims
45 to 47, wherein the IMPDH mediated disease is an
inflammatory disease and wherein said additional agent,
5 if present, is an antiinflammatory agent.
INTERNATIONAL SEARCH REPORT
Intei national Application No
PCT/US 97/06623
A. CLASSIFICATION OF SUBJECT MATTER
IPC 6 C07D263/32 A61K31/42 C07D413/12 C07C275/28
C07C275/42
According to international Patent Clarification (IPC) or to both national classification and IPC
C07C275/34
B. FIELDS SEARCHED
Minimum documentation searched (classification system followed by classification symbols)
IPC 6 C07D C07C
Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched
Electronic data base consulted during the international search (name of data base and, where practical, search terms used)
C. DOCUMENTS CONSIDERED TO BE RELEVANT
Category " Citation of document, with indication, where appropriate, of the relevant passages
Relevant to claim No.
US 5 380 879 A (ERIC B. SJOGREN ) 10
January 1995
cited in the application
see column 17-column 23
see column 2, line 20 - line 55
1-53
□
Further documents are listed m the continuation of box C.
Patent family members are Listed in annex.
* Special categories of cited documents :
"A* document defining the general state of the art which is not
considered to be of particular relevance
"E" earlier document but published on or after the international
filing date
"L" document which may throw doubts on priority claimfs) or
which is cited to establish the publication date of another
citation or other special reason (as specified)
"O" document referring to an oral disclosure, use, exhibition or
other means
"P" document published prior to the international filing date but
later than the priority date claimed
*T" later document published after the international filing date
or pnonty date and not in conflict with the application but
cited to understand the principle or theory underlying the
invention
'X' document of particular relevance; the claimed invention
cannot be considered novel or cannot be considered to
involve an inventive step when the document is taken alone
"Y" document of particular relevance; the claimed invention
cannot be considered to involve an inventive step when the
document is combined with one or more other such docu-
ments, such combination being obvious to a person skilled
in the art.
document member of the same patent family
Date of the actual completion of the international search
24 July 1997
Date of mailing of the international search report
0 4. 08. $7
Name and mailing address of the ISA
European Patent Office, P.B. 581 B Palentlaan 2
NL - 2280 HV Rijswijk
Tel. ( + 31-70) 340-2040, Tx. 31 651 epo nl.
Fax: ( + 31-70) 340-3016
Authorized officer
Henry, J
Form PCT/ISA.210 (second <heel) (July 1992)
INTERNATIONAL SEARCH REPORT
ernattonal application No.
PCT/US 97/06623
Box I Observations where certain claims were found unsearchable (Continuation of item 1 of first sheet)
This Inlernauona! Search Report has not been established in respect of certain claims under Article I7(2)(a) for the following reasons:
1. Claims Nos.:
because they relate to subject matter not required to be searched by this Authority, namely:
Remark: Although claim(s) 1-8, 45-53
is(are) directed to a method of treatment of the human/animal
body, the search has been carried out and based on the alleged
effects of the compound/composition.
because they relate to parts of the International Application that do not comply with the prescribed requirements to such
an extent that no meaningful InternauonaJ Search can be carried out, specifically:
In view of the large number of compounds which are defined by the wording
of the claims, the search has been performed on the general idea and
compounds mentioned in the examples of the description.
Claims searched incompletely: 1-53
because they are dependent claims and are not drafted in accordance with the second and third sentences of Rule 6.4(a).
Box II Observations where unity of invention is lacking (Continuation of item 2 of first sheet)
This InternauonaJ Searching Authority found multiple invenuons in this international application, as follows:
j | I As jji required additional search fees were timely paid by the applicant, this International Search Report covers all
I 1 searchable claims.
2 I I As searchable claims could be searched without efTort justifying an additional fee. this Authority did not invite payment
of any additional fee.
3 j 1 As only some of the required additional search fees were timely paid by the applicant, this International Search Report
I 1 covers only those claims for which fees were paid, specifically claims Nos.:
4 I I No required additional search fees were timely paid by the applicant. Consequently, this Inlernauonal Search Report is
I 1 restricted to the invenuon first mentioned in the claims; it is covered by claims Nos.:
Remark on Protest The additional search fees were accompanied by the applicant s protest.
i j No protest accompanied the payment of additional search fees.
p orm PCT,'ISA,'210 (continuation of first sheet (1)) (July 1992)
INTERNATIONAL SEARCH REPORT
Information on patent family members
Intcn.aOonal Application No
PCT/US 97/G6623
Patent document
cited in search report
Publication
dale
Patent family
member(s)
Publication
date
US 5380879 A
10-01-95
AU
CA
CN
EP
FI
WO
US
1916995
2183529
1143366
Q745074
963219
9522535
5441953
04-09-95
24-08-95
19-02-97
04-12-96
11-10-96
24-08-95
15-08-95
Form PCT,'ISA,ai0 (patent family annex) (July 1992)
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